Reducing the amount of error and introduced data variability increases the accuracy of Western blot results. In this study, different methods of normalization for loading differences and data alignment were explored with respect to their impact on Western blot results. GAPDH was compared to the LI-COR Revert total protein stain as a loading control. The impact of normalizing data to a control condition, which is commonly done to align Western blot data distributed over several immunoblots, was also investigated. Specifically, this study addressed whether normalization to a small subset of distinct controls on each immunoblot increases pooled data variability compared to a larger set of controls. Protein expression data for NOX-2 and SOD-2 from a study investigating the protective role of the bradykinin type 1 receptor in angiotensin-II induced left ventricle remodeling were used to address these questions but are also discussed in the context of the original study. The comparison of GAPDH and Revert total protein stain as a loading control was done by assessing their correlation and comparing how they affected protein expression results. Additionally, the impact of treatment on GAPDH was investigated. To assess how normalization to different combinations of controls influences data variability, protein data were normalized to the average of 5 controls, the average of 2 controls, or an average vehicle and the results by treatment were compared. The results of this study demonstrated that GAPDH expression is not affected by angiotensin-II or bradykinin type 1 receptor antagonist R-954 and is a less sensitive loading control compared to Revert total protein stain. Normalization to the average of 5 controls tended to reduce pooled data variability compared to 2 controls. Lastly, the results of this study provided preliminary evidence that R-954 does not alter the expression of NOX-2 or SOD-2 to an expression profile that would be expected to explain the protection it confers against Ang-II induced left ventricle remodeling.

Premature babies are at risk of death from immature lung development. For this reason, pregnant mothers at risk for preterm delivery are administered dexamethasone (DEX), a synthetic glucocorticoid that promotes fetal lung development. However, exposure to DEX in utero is associated with low birth weight and cardiovascular development pathologies. Moreover, our lab found that DEX administration in-utero leads to a sex-specific increase in stress-induced tachycardia in female, but not male offspring. This project seeks to expand on this preliminary finding of the heart by examining local effectors of activity from the sympathetic system (tyrosine hydroxylase and catechol-o-methyltransferase). Tyrosine hydroxylase was measured as it catalyzes the rate limiting step of norepinephrine synthesis while catechol-O- methyltransferase was studied as it catalyzes the degradation of norepinephrine. Acetylcholinesterase was used to measure parasympathetic activity as it catalyzes the degradation of the primary neurotransmitter of the parasympathetic nervous system, acetylcholine. Analyses of sympathetic as well as parasympathetic activity were done to determine influences of in-utero DEX exposure on autonomic regulation in adulthood. Pregnant rats were administered DEX (0.4 mg/kg, i.p.) or vehicle (20% w/v 2-hydroxypropyl ß- cyclodextran) at gestation days 18-21, with euthanasia of offspring occurring at around the time the offspring reached 13-15 weeks of age. Left ventricles and right atria were pulverized, processed and subjected to western blot analysis to determine expression of proteins of interest. Males exposed to DEX in-utero saw a decrease in tyrosine hydroxylase expression in left ventricle and right atrium when compared to vehicle control, a difference not seen with females. In addition, catechol-o-methyltransferase expression was increased in right atria from male, but not female rats. Acetylcholinesterase expression was reduced in the right atria of female, but not male rats. The present findings suggest reduced norepinephrine signaling in the heart of male, but not female DEX-exposed offspring. Given that we have previously found that female, but not male rats exhibit exaggerated stress-induced tachycardia, our current findings suggest that males possess a sex-specific compensatory mechanism allowing the heart to resist increased sympathetic signaling from the brain, one that females do not possess. The underlying mechanics of this proposed mechanism are unclear, and further investigation is needed in this subject to determine the significance of the findings from our study.

In the 1950s and 1960s, researchers Leon Chesley, John Annitto, and Robert Cosgrove investigated the possible familial factor for the conditions of preeclampsia and eclampsia in pregnant women. Preeclampsia and eclampsia, which are related to high blood pressure, have unknown causes and affect at least five percent of all pregnancies. The researchers, who worked at Margaret Hague Maternity Hospital in Jersey City, New Jersey, used hospital patient records to find and reexamine women who had eclampsia at the hospital, as well as their daughters, sisters, daughters-in-law, and granddaughters. Chesley and colleagues found that the daughters and granddaughters of eclamptic women were more likely than the female offspring of non-eclamptic women to have preeclampsia and eclampsia in their own pregnancies, and especially in their first pregnancies. The study provided evidence that the disorders are inherited, enabling physicians to better monitor pregnancies in women who have a known family history for preeclampsia and eclampsia.

Leon Chesley studied hypertension, or high blood pressure, in pregnant women during the mid-twentieth century. Chesley studied preeclampsia and eclampsia, two hypertensive disorders found in approximately five percent of all US pregnancies. In New Jersey and New York, Chesley devoted over forty years to researching preeclampsia and eclampsia. Chesley conducted several long-term studies using the same group of women beginning from their pregnancies. Chesley’s multi-decade research led to more accurate diagnosis of preeclampsia and eclampsia in pregnant women and significantly reduced the mortality of pregnant women due to hypertensive diseases.

Leon Chesley published Hypertensive Disorders in Pregnancy in 1978 to outline major and common complications that occur during pregnancy and manifest in abnormally high blood pressures in pregnant women. The book was published by Appleton-Century-Crofts in New York, New York. Chesley compiled his book as a tool for practicing obstetricians and teachers. The book focuses on preeclampsia and eclampsia, but it also describes other common and rare hypertensive diseases and disorders of pregnancy and discusses their histories, diagnoses, management plans, pathologies, and immediate and remote prognoses for mothers and fetuses. Doctors used the book and all subsequent editions to help diagnose and manage complications during pregnancy and to avoid deaths for pregnant women and fetuses.

In April 1994, Elizabeth Raymond, Sven Cnattingius, and John Kiely published “Effects of Maternal Age, Parity, and Smoking on the Risk of Stillbirth” in the British Journal of Obstetrics and Gynecology, now known as BJOG: An International Journal of Obstetrics and Gynecology. The article examines how advanced maternal age, defined as delivery at thirty-five years old or older, cigarette smoking, and nulliparity, or the state of never having given birth, can negatively impact pregnancy. At the time of publication, according to Raymond and colleagues, stillbirths comprised over half of all perinatal, or close to birth, deaths and more than one-third of total fetal and infant deaths in Europe and North America. In the article, Raymond and her coauthors demonstrate how certain risk factors may increase the risk of stillbirth at different stages of pregnancy, which helped set a foundation for future research in interventions to prevent stillbirth.