Chander Arora

Long-term nitric oxide blockade in the pregnant rat: Effects on blood pressure and plasma levels of endothelin-1

OBJECTIVE Abnormalities in the production of nitric oxide and endothelin-1 have been implicated in the development of preeclampsia. We postulated that long-term nitric oxide synthase inhibition with L-nitro-arginine methyl ester would induce sustained hypertension, a rise in plasma levels of endothelin-1, and fetal growth restriction. STUDY DESIGN Conscious virgin and pregnant Sprague-Dawley rats received infusions of vehicle or L-nitro-arginine methyl ester (2.5 mg/kg/hr) for 11 days. Mean arterial pressure was assessed serially. On day 21 of gestation (or equivalent in virgin rats) plasma was collected for endothelin-1 levels; pup weight and litter size were determined. Data were analyzed with analysis of variance and regression techniques. RESULTS Mean arterial pressure was constant in virgin control rats (n = 7) but declined in pregnant control rats (n = 11) as gestation advanced. Nitric oxide synthase inhibition in virgin (n = 10) and pregnant (n = 11) rats caused sustained elevations in mean arterial pressure (165 +/- 7 vs 100 +/- 3 mm Hg, L-nitro-arginine methyl ester vs control virgin rats, p < 0.0001; 149 +/- 5 vs 91 +/- 2 mm Hg, L-nitro-arginine methyl ester vs control pregnant rats, p < 0.0001). L-nitro-arginine methyl ester induced a rise in plasma endothelin-1 levels in virgin (4.4 +/- 0.1 vs 3.5 +/- 0.1 pg/ml, L-nitro-arginine methyl ester vs control, p < 0.0001) and pregnant rats (3.0 +/- 0.1 vs 2.6 +/- 0.1 pg/ml, L-nitro-arginine methyl ester vs control, p < 0.0001). Pregnant rats had lower endothelin-1 levels than did virgin rats (p < 0.0001). Mean arterial pressure and endothelin-1 were significantly correlated in pregnant rats. L-nitro-arginine methyl ester decreased pup weight (2.4 +/- 0.4 vs 3.7 +/- 0.2 gm/pup/litter, L-nitro-arginine methyl ester vs control, p < 0.01) and litter size (6.6 +/- 1.3 vs 10.2 +/- 0.9 pups/litter, L-nitro-arginine methyl ester vs control, p < 0.05). CONCLUSIONS Long-term nitric oxide synthase blockade causes sustained hypertension, elevated levels of endothelin-1, and fetal growth restriction. Although the endocrine and pressor effects are not unique to pregnancy, this model clearly induces some of the changes seen in preeclampsia and may be useful for studying specific interventions.

Biomarkers of Spontaneous Preterm Birth: An Overview of The Literature in the Last Four Decades

Background: Understanding spontaneous preterm birth ([PTB] < 37 weeks) is difficult due to heterogeneities associated with multitudes of risk factors and pathophysiological pathways. Several biomarkers are routinely used clinically for predicting preterm labor; however, these factors are either nonspecific or detected too late. Objective: Systematic review of literature on PTB biomarkers in the last 40 years to map out the existing knowledge and gaps in understanding PTB biomarkers. Search strategies: Five electronic databases were searched for human studies on PTB biomarkers published in any language between 1965 and 2008. Selection criteria: The phenotype of interest for final data extraction was exclusively spontaneous PTB with no rupture of membranes. Data extraction included (a) general characteristics of the study (clinical setting, period, and study design), (b) study/participant characteristics (inclusion and exclusion criteria, race/ethnicity, number of participants, gestational age at sampling, (c) characteristics of the biomarker (type, rationale for its selection, type of biological sample, and assay used, and (d) concentration of biomarkers in cases and controls. Data collection and analysis: The search yielded 7255 citations and data were extracted from 217 articles which met our inclusion and exclusion criteria. Main results: A total of 116 different biomarkers were reported and these were assayed 578 times in the 217 included studies. Over two thirds of the 217 studies were performed on North American or European populations. No reliable biomarkers emerged as a risk predictor of PTB. Conclusions: Identifying similar studies on biomarkers for the prediction of PTB was a very challenging task due heterogeneities in study design, sampling issues (types, timing and processing), assay methods, and analyses. Major areas of concern identified in this review include poor phenotype definition, nonideal study designs and poor rationale for biomarker selection and assays and population stratification issues.

Corticotrophin-releasing hormone and CRH-binding protein. Differences between patients at risk for preterm birth and hypertension.

Abstract: Background: During pregnancy in the second and third trimester there is a progressive rise in plasma CRH thought to be secreted by the placenta. Plasma CRH‐BP inactivates CRH, which may prevent its peripheral action on the maternal pituitary and myometrium. In the last few weeks of pregnancy CRH‐BP decreases, thereby causing an increase in free CRH or a CRH/CRH‐BP complex available to play a role in the onset of parturition. Objective: We tested the hypothesis that differences in CRH, CRH‐BP, or a CRH/CRH‐BP complex in patients at risk for preterm birth (PTB) and hypertension (HYP) account for the differences in the timing of parturition. Methods: From a Behavior in Pregnancy Study database, we identified 18 patients who had spontaneous PTB and 23 patients who developed HYP. Both groups were case controlled and matched with patients who delivered at term (Normal). Maternal plasma samples had been appropriately collected from these patients at 18‐20, 28‐30, and 35‐36 weeks gestational age. CRH levels were measured by double antibody RIA kit and the CRH‐BP by a immunoradiometric technique. A CRH‐BP/CRH dimer complex index was calculated. Statistical analysis was done using Kruskal‐Wallis test for two cases. Results: Maternal CRH (pg/ml) in the PTB cases compared to the HYP cases was significantly elevated at all three time periods. Maternal CRH‐BP (pg/ml) in the PTB versus HYP cases was significantly lower at all three time periods in the PTB cases compared to the HYP cases. Maternal CRH‐BP/CRH dimer complex index was significantly lower in the PTB cases at all three time periods than either the controls or the HYP cases, suggesting excessive CRH. The mean GA at delivery for the PTB cases was significantly lower than the control or HYP cases. Conclusions: These results suggest that those patients at risk for PTB have significantly elevated CRH, lower CRH‐BP, and a reduced CRH‐BP/CRH dimer complex index at all three time periods of assessment.

Placental Corticotropin-Releasing Hormone Mediates the Association Between Prenatal Social Support and Postpartum Depression

Three decades of research point to both biological and psychological risk factors for postpartum depression, but very little research integrates the two. This study bridged this gap by testing whether prenatal social support predicted depressive symptoms at 8 weeks postpartum in a multiethnic sample of 210 women and whether the stress hormone placental corticotropin-releasing hormone (pCRH), measured at 19, 29, and 37 weeks’ gestation, mediated this relationship. We found that prenatal family support predicted significantly fewer depressive symptoms postpartum and more gradual increases in pCRH from 29 to 37 weeks’ gestation. Furthermore, steeper increases in pCRH during this same period predicted more depressive symptoms postpartum. Finally, these changes in pCRH in late pregnancy mediated the relationship between prenatal family support and postpartum depressive symptoms. These results suggest that social and biological risk factors for postpartum depressive symptoms are intertwined and move us closer to an integrated biopsychosocial understanding of postpartum depression.

Plasma atrial natriuretic peptide levels during the rat estrous cycle, pregnancy, and puerperium

The rat has been used as a model for studying the changes that occur in maternal blood volume and renal function during pregnancy. The role, if any, that atrial natriuretic peptide plays in regulating these changes is unknown, and little information is available on atrial natriuretic peptide levels at different stages of gestation in the rat. In this study we measured plasma atrial natriuretic peptide levels by radioimmunoassay in the rat at each stage of the estrous cycle, during the last 2 weeks of pregnancy, and in the early postpartum period. Atrial natriuretic peptide levels did not change during the estrous cycle. Atrial natriuretic peptide levels were low on days 10 to 15 of gestation but rose to become significantly higher than nonpregnant levels on days 16 to 18. On day 21 shortly before delivery, levels were similar to nonpregnant values. Postpartum, atrial natriuretic peptide levels rose immediately and remained elevated for the next 48 hours. These findings suggest that factors other than blood volume may mediate plasma atrial natriuretic peptide levels during pregnancy and the postpartum period.

Prenatal Air Pollution Exposure and Ultrasound Measures of Fetal Growth in Los Angeles, California

BACKGROUND Few previous studies examined the impact of prenatal air pollution exposures on fetal development based on ultrasound measures during pregnancy. METHODS In a prospective birth cohort of more than 500 women followed during 1993-1996 in Los Angeles, California, we examined how air pollution impacts fetal growth during pregnancy. Exposure to traffic related air pollution was estimated using CALINE4 air dispersion modeling for nitrogen oxides (NOx) and a land use regression (LUR) model for nitrogen monoxide (NO), nitrogen dioxide (NO2) and NOx. Exposures to carbon monoxide (CO), NO2, ozone (O3) and particles <10μm in aerodynamic diameter (PM10) were estimated using government monitoring data. We employed a linear mixed effects model to estimate changes in fetal size at approximately 19, 29 and 37 weeks gestation based on ultrasound. RESULTS Exposure to traffic-derived air pollution during 29 to 37 weeks was negatively associated with biparietal diameter at 37 weeks gestation. For each interquartile range (IQR) increase in LUR-based estimates of NO, NO2 and NOx, or freeway CALINE4 NOx we estimated a reduction in biparietal diameter of 0.2-0.3mm. For women residing within 5km of a monitoring station, we estimated biparietal diameter reductions of 0.9-1.0mm per IQR increase in CO and NO2. Effect estimates were robust to adjustment for a number of potential confounders. We did not observe consistent patterns for other growth endpoints we examined. CONCLUSIONS Prenatal exposure to traffic-derived pollution was negatively associated with fetal head size measured as biparietal diameter in late pregnancy.

Perinatal factors influencing atrial natriuretic peptide levels in umbilical arterial plasma at the time of delivery

Little is known about atrial natriuretic peptide metabolism or secretion in the human fetus. The purpose of this study was to determine if both the placenta and umbilical vessels are possible sites of atrial natriuretic peptide metabolism and to evaluate the effects that labor, route of delivery, prolonged pregnancy, preeclampsia, and fetal distress have on umbilical arterial atrial natriuretic peptide levels. We found that plasma atrial natriuretic peptide levels in the umbilical artery are significantly greater than those in the vein (p less than 0.001). Umbilical arterial and umbilical venous atrial natriuretic peptide levels were higher in plasma samples collected immediately at delivery when compared with those obtained 10 minutes later (p less than 0.001). Umbilical arterial atrial natriuretic peptide levels were elevated in pregnancies complicated by preeclampsia and fetal distress (p less than 0.01). Labor, route of delivery, and prolonged pregnancy had no effect on umbilical arterial atrial natriuretic peptide levels. We propose that both the placenta and umbilical vessels contain atrial natriuretic peptide receptors that are involved in the clearance or metabolism of atrial natriuretic peptide. The increased umbilical arterial atrial natriuretic peptide levels present in preeclampsia and fetal distress may reflect an attempt by the fetus to regulate blood flow.

Corticotrophin-Releasing Hormone in Lipopolysaccharide-Stimulated Term Fetal Membranes and Amniotic Fluid From Term and Preterm Birth in African Americans and Caucasians

The objective of this study is to document differences in corticotrophin-releasing hormone (CRH), CRH receptor 1 (CRHR1), and CRH binding protein (CRHBP) gene expression in fetal membranes derived from African Americans and Caucasians in vitro in response to lipopolysaccharide (LPS) stimulation and to assess racial disparity in CRH concentrations in the amniotic fluid (AF) of women with spontaneous preterm birth (PTB). Fetal membranes (African American, n = 8; Caucasian, n = 8) at term, placed in an organ explant system, were stimulated with LPS. Microarray analysis documented differences in the mRNA expression pattern of CRH, CRHBP, and CRHR1 between races. CRH was measured in AF (a case [PTB]—control [term] study) and culture media. Between races, LPS significantly increased CRH and CRHR1 expression in African Americans and CRHBP in Caucasians, with no differences in controls. CRH was detectable only in LPS-stimulated African American membranes. AF CRH concentrations were higher in PTB compared with controls (P < .001), and no difference was noticed between races (P = .1). AF analysis did not document racial disparity in CRH concentrations in PTB. In fetal membranes, African Americans showed a higher expression and production of CRH in response to an in vitro stimulus.

Atrial natriuretic peptide metabolism during pregnancy in the rat.

OBJECTIVE Our purpose was to determine whether plasma clearance rates and production rates of atrial natriuretic peptide 99-126 are altered during pregnancy in the rat. STUDY DESIGN Twelve virgin and 12 late-pregnant chronically instrumented, conscious, unrestrained Sprague-Dawley rats were studied. Mean arterial pressure, heart rate, and plasma atrial natriuretic peptide levels were measured before and during a 40-minute continuous infusion of atrial natriuretic peptide (10 ng/kg/min). RESULTS Control mean arterial pressure was 106 +/- 5 mm Hg in virgin rats versus 97 +/- 4 mm Hg in pregnant rats. Atrial natriuretic peptide infusion did not significantly affect mean arterial pressure in either group of animals but decreased heart rate in virgin rats. Basal plasma atrial natriuretic peptide levels were significantly higher in virgin than in pregnant rats (107 +/- 10 vs 78 +/- 7 pg/ml, respectively, p < 0.05). Atrial natriuretic peptide infusion significantly increased plasma levels in both groups to similar (183 +/- 19 and 154 +/- 14 pg/ml, virgin vs pregnant rats). Calculated plasma clearance rates were similar in virgin and pregnant rats (166 +/- 27 vs 155 +/- 17 ml/kg/min). Estimated production rates of atrial natriuretic peptide were higher in virgin then in pregnant rats (15.1 +/- 1.4 vs 11.4 +/- 1.1 ng/kg/min, p < 0.05). CONCLUSIONS Plasma atrial natriuretic peptide levels are lower in chronically instrumented near-term pregnant rats compared with levels in virgin rats. This is not related to differences in plasma atrial natriuretic peptide clearance rates but rather to a decrease in production rates in late pregnancy.

Effect of pregnancy on the vasodepressor response to atrial natriuretic factor

OBJECTIVE We attempted to determine whether pregnancy alters the vasodepressor response to both physiologic and pharmacologic infusions of atrial natriuretic factor 99-126. STUDY DESIGN Ten virgin and 10 pregnant (17 +/- 1 days of gestation) conscious, unrestrained Sprague-Dawley rats with chronic indwelling vascular catheters were studied. Mean arterial pressure and heart rate were measured in response to steady-state infusions of either saline solution or increasing concentrations of atrial natriuretic factor (range 5 to 2560 ng.kg-1.min-1). RESULTS Basal mean arterial pressure was significantly lower in pregnant rats than in virgin rats (89 +/- 3 vs 97 +/- 2 mm Hg, p < 0.02). Atrial natriuretic factor induced significant dose-dependent decreases in mean arterial pressure and heart rate in virgin and pregnant rats (p < 0.001). The hypotensive effects of atrial natriuretic factor were blunted in the pregnant rats only in response to the highest concentrations of atrial natriuretic factor administered (-27 +/- 3 mm Hg in pregnant rats vs -43 +/- 3 mm Hg in virgin rats, p < 0.005). CONCLUSIONS The vasodepressor response to physiologic infusions of atrial natriuretic factor was not affected by pregnancy status. However, pharmacologic infusions of atrial natriuretic factor resulted in a blunted vasodepressor response in the pregnant animals. This may be due to alterations in vascular atrial natriuretic factor receptors, changes in the clearance rate of atrial natriuretic factor, or the modulating effects of other vasoactive hormones.