ABSTRACT
INTRODUCTION: The effective management of pre-eclampsia (PE), a complex condition affecting 5% of pregnancies [1], would benefit from a clear diagnostic assay. Due to this need, great interest lies in the development and validation of objective biomarkers, among which antibodies remain attractive given their amplification by the immune system, stability, and current clinical use. Although several recent studies lend support to the idea that autoantibodies against the angiotensin II AT1 receptor (AT1-AAs) could be involved in PE [1], their presence is not highly specific for PE as measured by cell-based assays for AT1 agonism [2]. Given these results, we hypothesized that additional antibody biomarkers may exist in PE, and that a small panel of such biomarkers could provide an effective diagnostic tool. Furthermore, development of a simple binding assay will facilitate AT1-AA detection for larger-scale studies, while identifying other antibody biomarker(s) would enhance understanding of the immune component to PE pathogenesis. OBJECTIVES: The primary objective of this study was to identify a panel of peptide reagents that preferentially bind PE patients' serum antibodies. In addition, we sought to measure the frequency of AT1-AAs in a new patient cohort using a simplified assay. METHODS: Plasma samples were obtained from normal-outcome pregnancies (n=28) as well as PE patients (n=25) and enriched for the antibody fraction. The seven amino acid AT1-AA epitope [1] was expressed on the surface of E. coli for reactivity analysis by flow cytometry. A bacteria-displayed linear peptide library was screened against the antibody repertoires using a unique method of fluorescence activated cell sorting (FACS) to isolate peptides that react with multiple patients and show little reactivity with normal-outcome pregnancies. FACS analysis was used to measure individual peptide reactivity, and statistical analyses included a Student's t-test and nonparametric tests to compare the means, medians, and normal scores. RESULTS: Using this simple binding assay, AT1-AAs were detected in a majority of PE patients and more rarely in normal-outcome pregnancies. Among the isolated peptide mimics, several unique peptides demonstrated significantly (p<0.03) higher reactivity with PE patients than with control samples but showed no sequence homology to the known AT1 epitope. Not only did our peptides perform well with the original set of samples used for discovery, but these peptides also reacted with antibodies from a small set of new PE (n=10) sera but not from new normal-outcome (n=10) pregnancies. CONCLUSION: Peptides distinct from the AT1 epitope and identified by library screening exhibited potential diagnostic utility for PE, suggesting that a panel of such peptides might provide a novel diagnostic test to distinguish PE from other conditions with similar symptoms. Also, this study demonstrated AT1-AA presence in an independent patient cohort with a simple binding assay, which can be easily expanded to evaluate larger cohorts.
ABSTRACT
To determine the effect of cystocele upon voiding, 30 women with various degrees of genitourinary prolapse were studied. The patients were divided into three groups depending on the severity of the cystocele and were evaluated with uroflowmetry, urethrocystoscopy, water cystometry, urethral pressure profilometry and voiding urethrocystometry. The three groups were similar in most parameters except maximum urethral closure pressure (P less than .05). The patients with cystocele did not demonstrate the abnormal voiding patterns characteristic of outflow obstruction.
Subject(s)
Urinary Bladder Diseases/physiopathology , Urination , Urodynamics , Adult , Aged , Female , Humans , Middle AgedABSTRACT
Fetal heart rate tracings of pregnancies complicated by cocaine use were analyzed to evaluate the effects of subacute maternal cocaine use on the fetus. Nonstress tests were performed twice weekly on patients from the perinatal substance abuse clinic with screening of maternal urine samples for cocaine, amphetamines, phencyclidine, and opiates at the time of each examination. Nonstress tests performed when the urine toxicology screen was positive for cocaine alone (positive cocaine nonstress tests) were paired with those from the same patient but performed when the screen was negative (negative cocaine nonstress tests). The nonstress tests were analyzed with the Lyons scoring system, which evaluates the baseline heart rate, the oscillatory amplitude of the baseline, the oscillatory frequency, decelerations, and accelerations. Twenty pairs of nonstress tests from 20 patients were analyzed. The total score was higher on the negative cocaine nonstress test in 70% of the pairs and equal in the remaining 30% (p less than 0.001). Significant differences occurred in the oscillatory amplitude (p less than 0.001), frequency (p = 0.002), and acceleration scores (p = 0.03) but not the fetal heart rate baseline or the deceleration scores. The observed changes may reflect alterations in fetal central nervous system neurotransmitters and fetal state regulation, which may affect the developing central nervous system of cocaine-exposed fetuses and in turn play a role in the developmental and behavioral abnormalities observed in cocaine-exposed infants.
