Pediatrics & Women's Health
Permanent URI for this collectionhttps://hdl.handle.net/20.500.12503/30293
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Browsing Pediatrics & Women's Health by Author "Cushen, Spencer"
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Item Absolute Quantification of Mitochondrial DNA in Peripheral Blood from Women with Preeclampsia(2020) Silzer, Talisa; Phillips, Nicole; Goulopoulou, Styliani; Reid, Danielle; Sun, Jie; Scroggins, Sabrina; Santillan, Mark; Santillan, Donna; Cushen, SpencerIntroduction Mitochondrial DNA (mtDNA) in maternal blood has been proposed as a potential predictor of preeclampsia (PE). The objective of this study was to use an absolute PCR (abPCR) quantification protocol to determine concentrations of mtDNA in maternal plasma and peripheral blood mononuclear cells (PBMC) from normal and PE pregnancies. Methods Blood samples were collected from pregnant women with uncomplicated pregnancies and pregnancies with PE (University of Iowa IRB#200910784). abPCR quantification of mtDNA and nDNA was performed on DNA extracts from plasma (in the presence or absence of lysis buffer) and PBMCs using TaqMan(TM) probes and chemistry. Results When plasma DNA was extracted using lysis buffer, mtDNA concentrations were lower in women with PE than in controls (Control: 4.83 ± 1.09 vs. PE: 1.72 ± 0.38 pg/uL, n=19, P=0.017), while concentrations of nDNA did not differ (P=0.39). Without lysis buffer, plasma mtDNA remained lower in women with PE compared to controls (Control: 0.0106 ± 0.0019 vs PE: 0.0019 ± 0.0003 pg/uL, n=16-20, P< 0.0001). There were no group differences in PBMC mtDNA (P=0.66) and nDNA (P=0.13) concentrations. Conclusion mtDNA concentrations were lower in plasma of pregnant women with PE compared to controls. A significant amount of mtDNA was membrane bound as indicated by a 480-fold greater concentration in DNA isolated from plasma with lysis buffer vs. without. Use of this improved method of quantification of mtDNA in multiple blood fractions may allow for its development as a biomarker to detect PE prior to the onset of organ damage.Item Postpartum Maternal Vascular Function in a Rat Model of Gestational Obstructive Sleep Apnea(2020) Osikoya, Oluwatobiloba; Wilson, Elizabeth; Mabry, Steve; Cushen, Spencer; Cunningham, Rebecca; Goulopoulou, Styliani; Singhal, JuhiIntroduction: De novo obstructive sleep apnea (OSA) in pregnancy is associated with adverse gestational outcomes. In pregnant mice, exposure to chronic intermittent hypoxia (CIH), a model of OSA, induces endothelial dysfunction in maternal uterine arteries. It is currently unknown whether gestational OSA has a long-term effect on maternal vascular function. We hypothesized that exposure to gestational CIH during pregnancy will result in postpartum maternal vascular dysfunction. Methods: Pregnant rats were assigned to Normoxia and CIH groups. The CIH group was exposed to five days of intermittent hypoxia [6 min cycles of 3 min hypoxia (10% O2) and 3 min normoxia (21% O2)]. Endothelial function was assessed in isolated maternal uterine arteries at weaning (postnatal day 28). Vascular reactivity to acetylcholine (ACh) was examined in the presence and absence of uterine perivascular adipose tissue (PVAT) using wire myography. Results: In the absence of PVAT, uterine arteries from dams exposed to gestational CIH had exaggerated responses to ACh compared to dams exposed to normoxia [(-)PVAT/pEC50, Normoxia: 6.77±0.08 vs. CIH: 7.13±0.09, p< 0.01], while PVAT normalized this difference [(+)PVAT/pEC50, Normoxia: 6.67±0.08 vs. CIH: 6.70±0.07, p=0.99]. The effects of PVAT were due to its anti-dilatory influences on uterine arteries from CIH-treated rats ([CIH (-PVAT) vs. CIH (+PVAT), p = 0.003), whereas it had no effect on arteries from normoxic rats (p = 0.77). Conclusion: Exposure to gestational CIH exaggerated postpartum uterine vascular smooth muscle relaxation responses. Gestational OSA may impair maternal vascular recovery after birth.