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1.
J Neonatal Perinatal Med ; 11(3): 305-310, 2018.
Article in English | MEDLINE | ID: mdl-30198877

ABSTRACT

OBJECTIVE: Pregnant non-Hispanic blacks (NHB) have increased vaginal microbiome diversity compared to non-Hispanic whites (NHW) which may contribute to increased preterm birth. Cervical microbiome diversity is poorly characterized in pregnancy, therefore our objective was to correlate cervical microbiota diversity with cervico-vaginal inflammation by race and delivery timing. STUDY DESIGN: Pregnant women were recruited in the first and second trimesters. A sterile cervical swab and saline lavage were collected at a single time point. Using 16S rRNA sequencing, Chao1 and Shannon Diversity (SDI) indicies were measured and compared by race and delivery timing (preterm vs. term delivery). Cervico-vaginal inflammatory markers were also compared by race and delivery timing. Spearman correlation coefficients between cervical microbiome diversity and cervico-vaginal inflammatory markers were calculated. RESULTS: Of the 51 subjects, 39 (76%) were NHB and 12 (24%) were NHW. Cervical microbiota SDI was significantly higher in NHB compared to NHW (0.5 vs. 0.1; p = 0.03). However, there were no difference in Chao1 diversity or cervico-vaginal inflammatory markers by race or delivery timing. CONCLUSION: Our findings suggest the cervical microbiota diversity during pregnancy differs by race. Larger cohort studies will further determine if altered cervical diversity is part of the pathogenesis of PTB and explains race disparities.


Subject(s)
Black or African American , Cervix Uteri/microbiology , Microbiota/physiology , Premature Birth/epidemiology , RNA, Ribosomal, 16S/physiology , Vagina/microbiology , White People , Adult , Cervix Uteri/physiology , Female , Humans , Infant, Newborn , Pregnancy , Pregnancy Outcome , Premature Birth/ethnology , Prospective Studies , Sequence Analysis, RNA , United States/epidemiology , Vagina/physiology
2.
Biol Reprod ; 94(5): 119, 2016 05.
Article in English | MEDLINE | ID: mdl-27009041

ABSTRACT

Preterm premature rupture of membranes (PPROM) is often associated with intra-amniotic inflammation and infection. Current understanding of the pathogenesis of PPROM includes activation of pro-inflammatory cytokines and proteolytic enzymes leading to compromise of membrane integrity. The impact of exposure to bacterial pathogens, including Ureaplasma parvum, on gestational membranes is poorly understood. Our objective was to develop a dual-chamber system to characterize the inflammatory response of gestational membranes to U. parvum in a directional nature. Full-thickness human gestational membrane explants, with either choriodecidua or amnion oriented superiorly, were suspended between two washers in a cylindrical device, creating two distinct compartments. Brilliant green dye was introduced into the top chamber to assess the integrity of the system. Tissue viability was evaluated after 72 h using a colorimetric cell proliferation assay. Choriodecidua or amnion was exposed to three doses of U. parvum and incubated for 24 h. Following treatment, media from each compartment were used for quantification of U. parvum (quantitative PCR), interleukin (IL)-8 (enzyme-linked immunosorbent assay), and matrix metalloproteinase (MMP)-2 and MMP-9 activity (zymography). We observed that system integrity and explant viability were maintained over 72 h. Dose-dependent increases in recovered U. parvum, IL-8 concentration, and MMP-2 activity were detected in both compartments. Significant differences in IL-8 concentration and MMP-9 activity were found between the choriodecidua and amnion. This tissue explant system can be used to investigate the inflammatory consequences of directional bacterial exposure for gestational membranes and provides insight into the pathogenesis of PPROM and infectious complications of pregnancy.


Subject(s)
Chorioamnionitis/microbiology , Chorioamnionitis/pathology , Extraembryonic Membranes/pathology , Pregnancy Complications, Infectious/pathology , Tissue Culture Techniques/methods , Ureaplasma Infections/pathology , Ureaplasma/physiology , Amnion/metabolism , Chorioamnionitis/metabolism , Cytokines/metabolism , Extraembryonic Membranes/metabolism , Female , Fetal Membranes, Premature Rupture/metabolism , Fetal Membranes, Premature Rupture/pathology , Humans , Inflammation Mediators/metabolism , Models, Biological , Pregnancy , Pregnancy Complications, Infectious/metabolism , Pregnancy Complications, Infectious/microbiology , Tissue Culture Techniques/instrumentation , Ureaplasma/isolation & purification , Ureaplasma Infections/metabolism
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