Four‐Dimensional Ultrasonography of the Fetal Heart Using Color Doppler Spatiotemporal Image Correlation

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/135517/1/jum2004234473.pd

miR-143 Regulation of Prostaglandin-Endoperoxidase Synthase 2 in the Amnion: Implications for Human Parturition at Term

The human amnion plays a pivotal role in parturition. Two of its compartments, the placental amnion and the reflected amnion, have distinct transcriptome and are functionally coordinated for parturition. This study was conducted to determine the microRNA (miRNA) expression pattern and its significance in the placental amnion and the reflected amnion in association with labor at term.<0.05). Luciferase assay and transfection confirmed miR-143 binding to 3′ UTR of prostaglandin-endoperoxidase synthase 2 (PTGS2) mRNA and miR-143 regulation of PTGS2 in AMCs.We report region-specific amniotic microRNAome and miR-143 regulation of PTGS2 in the context of human labor at term for the first time. The findings indicate that miRNA-mediated post-transcriptional regulation of gene expression machinery in the amnion plays an important role in the compartments (placental amnion vs reflected amnion) and in a cell type-specific manner (AECs vs AMCs) for parturition

Human βâ defensinâ 1: A natural antimicrobial peptide present in amniotic fluid that is increased in spontaneous preterm labor with intraâ amniotic infection

ProblemHuman βâ defensins (HBDs) are antimicrobial peptides that participate in the soluble innate immune mechanisms against infection. Herein, we determined whether HBDâ 1 was present in amniotic fluid during normal pregnancy and whether its concentrations change with intraâ amniotic inflammation and/or infection.Method of StudyAmniotic fluid was collected from 219 women in the following groups: (a) midtrimester who delivered at term (n = 35); (b) term with (n = 33) or without (n = 17) labor; (c) preterm labor with intact membranes who delivered at term (n = 29) or who delivered preterm with (n = 19) and without (n = 29) intraâ amniotic inflammation and infection or with intraâ amniotic inflammation but without infection (n = 21); and (d) preterm prelabor rupture of membranes (pPROM) with (n = 19) and without (n = 17) intraâ amniotic inflammation/infection. Amniotic fluid HBDâ 1 concentrations were determined using a sensitive and specific ELISA kit.Results(a) HBDâ 1 was detectable in all amniotic fluid samples; (b) amniotic fluid concentrations of HBDâ 1 were changed with gestational age (midtrimester vs term no labor), being higher in midtrimester; (c) amniotic fluid concentrations of HBDâ 1 were similar between women with and without spontaneous labor at term; (d) among patients with spontaneous preterm labor, amniotic fluid concentrations of HBDâ 1 in women with intraâ amniotic inflammation/infection and in those with intraâ amniotic inflammation without infection were greater than in women without intraâ amniotic inflammation or infection who delivered preterm or at term; and (e) the presence of intraâ amniotic inflammation and infection in patients with pPROM did not change amniotic fluid concentrations of HBDâ 1.ConclusionHBDâ 1 is a physiological constituent of amniotic fluid that is increased in midtrimester during normal pregnancy and in the presence of culturable microorganisms in the amniotic cavity. These findings provide insight into the soluble host defense mechanisms against intraâ amniotic infection.Amniotic fluid concentrations of human beta defensinâ 1 (HBDâ 1) in women with spontaneous preterm labor and intact membranes. Red lines indicate medians with interquartile ranges.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/146360/1/aji13031.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146360/2/aji13031_am.pd

Lipidomic analysis of patients with microbial invasion of the amniotic cavity reveals upâ regulation of leukotriene B4

Bioactive lipids derived from the metabolism of polyunsaturated fatty acids are important mediators of the inflammatory response. Labor per se is considered a sterile inflammatory process. Intraâ amniotic inflammation (IAI) due to microorganisms (i.e., intraâ amniotic infection) or danger signals (i.e., sterile IAI) has been implicated in the pathogenesis of preterm labor and clinical chorioamnionitis at term. Early and accurate diagnosis of microbial invasion of the amniotic cavity (MIAC) requires analysis of amniotic fluid (AF). It is possible that IAI caused by microorganisms is associated with a stereotypic lipidomic profile, and that analysis of AF may help in the identification of patients with this condition. To test this hypothesis, we analyzed the fatty acyl lipidome of AF by liquid chromatographyâ mass spectrometry from patients in spontaneous labor at term and preterm gestations. We report that the AF concentrations of proinflammatory lipid mediators of the 5â lipoxygenase pathway are significantly higher in MIAC than in cases of sterile IAI. These results suggest that the concentrations of 5â lipoxygenase metabolites of arachidonic acid, 5â hydroxyeicosatetraenoic acid, and leukotriene B4 in particular could serve as potential biomarkers of MIAC. This finding could have important implications for the rapid identification of patients who may benefit from antimicrobial treatment.â Maddipati, K. R., Romero, R., Chaiworapongsa, T., Chaemsaithong, P., Zhou, S.â L., Xu, Z., Tarca, A. L., Kusanovic, J. P., Gomez, R., Chaiyasit, N., Honn, K. V. Lipidomic analysis of patients with microbial invasion of the amniotic cavity reveals upâ regulation of leukotriene B4. FASEBJ. 30, 3296â 3307 (2016). www.fasebj.orgPeer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/154488/1/fsb2fasebj30100583.pd

Prevalence and Clinical Significance of Sterile Intra‐amniotic Inflammation in Patients with Preterm Labor and Intact Membranes

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/109340/1/aji12296.pd

A Novel Molecular Microbiologic Technique for the Rapid Diagnosis of Microbial Invasion of the Amniotic Cavity and Intra‐Amniotic Infection in Preterm Labor with Intact Membranes

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/106726/1/aji12189.pd

A Role for the Inflammasome in Spontaneous Labor at Term

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/143689/1/aji12440.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/143689/2/aji12440_am.pd

Identification of novel functional sequence variants in the gene for peptidase inhibitor 3

BACKGROUND: Peptidase inhibitor 3 (PI3) inhibits neutrophil elastase and proteinase-3, and has a potential role in skin and lung diseases as well as in cancer. Genome-wide expression profiling of chorioamniotic membranes revealed decreased expression of PI3 in women with preterm premature rupture of membranes. To elucidate the molecular mechanisms contributing to the decreased expression in amniotic membranes, the PI3 gene was searched for sequence variations and the functional significance of the identified promoter variants was studied. METHODS: Single nucleotide polymorphisms (SNPs) were identified by direct sequencing of PCR products spanning a region from 1,173 bp upstream to 1,266 bp downstream of the translation start site. Fourteen SNPs were genotyped from 112 and nine SNPs from 24 unrelated individuals. Putative transcription factor binding sites as detected by in silico search were verified by electrophoretic mobility shift assay (EMSA) using nuclear extract from Hela and amnion cell nuclear extract. Deviation from Hardy-Weinberg equilibrium (HWE) was tested by χ(2 )goodness-of-fit test. Haplotypes were estimated using expectation maximization (EM) algorithm. RESULTS: Twenty-three sequence variations were identified by direct sequencing of polymerase chain reaction (PCR) products covering 2,439 nt of the PI3 gene (-1,173 nt of promoter sequences and all three exons). Analysis of 112 unrelated individuals showed that 20 variants had minor allele frequencies (MAF) ranging from 0.02 to 0.46 representing "true polymorphisms", while three had MAF ≤ 0.01. Eleven variants were in the promoter region; several putative transcription factor binding sites were found at these sites by database searches. Differential binding of transcription factors was demonstrated at two polymorphic sites by electrophoretic mobility shift assays, both in amniotic and HeLa cell nuclear extracts. Differential binding of the transcription factor GATA1 at -689C>G site was confirmed by a supershift. CONCLUSION: The promoter sequences of PI3 have a high degree of variability. Functional promoter variants provide a possible mechanism for explaining the differences in PI3 mRNA expression levels in the chorioamniotic membranes, and are also likely to be useful in elucidating the role of PI3 in other diseases

Integrated Systems Biology Approach Identifies Novel Maternal and Placental Pathways of Preeclampsia

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