Open Tubular Capillary Electrochromatography-Laser Induced Fluorescence for the Separation and Detection of Proteins and Amino Acids

With its high efficiency and reproducibility, open tubular capillary electrochromatography (OTCEC) is beginning to prove itself as a promising method for the separation of proteins, peptides and pharmaceuticals. OTCEC, when coupled to an absorbance-based detector, suffers from poor sensitivity and high concentration limits of detection. However, laser induced fluorescence coupled to CEC has shown picomolar limits of detections for some compounds.2 This thesis describes the coupling of OTCEC to a laser induced fluorescence (LIF) detector to optimize both the separation and detection aspects of protein and amino acid analysis. OTCEC capillaries were etched and silanized, and the 4,4' cyanopentoxy biphenyl phase was attached through a radical-initiated process. Such biphenyl phases have shown unique dependence on temperature and mobile phase composition and demonstrated higher selectivity for certain compounds3 than bare silica capillaries. Proteins and amino acids were modified using pre-column derivatization formats with two different fluorescein-based dyes (fluorescein isothiocyanate, FITC) and Atto-tag FQ (3-(2-furoyl)quinoline-2-carboxaldehyde). These conditions were varied to attempt consistent labeling conditions with varying protein size. Optimizations of the analyses were achieved by modifying conditions such as background electrolyte, pH, voltage, and percent organic modifiers. These modified conditions were used to achieve the highest resolution on protein and amino acid derivatives. Results determined that high sensitivity and efficiency are achieved when LIF detection is coupled to cyano pentoxy biphenyl coated capillaries using optimal conditions. These conditions include 1) the use of Atto-tag FQ derivatization and 2) an 80:20 acetonitrile: 50 mM phosphate buffer (pH 2.00) background electrolyte with a positive electrical current applied

Introducing e-consents in a clinical setting

Preeclampsia (PE) is a multiorgan hypertensive-disorder in pregnancy that causes significant maternal-fetal mortality and morbidity. The diagnostics and therapeutics for PE are limited due to its unclear etiology. Using the UI Maternal Fetal Tissue Bank, our lab has demonstrated that copeptin is robustly predictive of PE. To investigate copeptin further, we developed the Rule Out Pre-Eclampsia Study (ROPE). The ROPE study recruits women admitted to Labor and Delivery for evaluation of PE. Women are admitted for PE evaluation at all hours. Research team members are not available at all times to obtain consent which limits recruitment. Our project aimed to develop an electronic informed consent (e-IC) that is compliant with the Federal Regulation for Human Research Protection and is easy to use and readily understood by study participants. After obtaining IRB approval, simulated patients were given an iPad on which to read and evaluate the e-IC using a validated questionnaire, the Quality of Informed Consent (QuIC). Based on the QuIC, the e-IC was modified and re-tested. Participants demonstrated good comprehension of the e-IC as evidence by QuIC scores ranging from 61 to 96. Based on our results, the e-IC is an effective and efficient method for the Informed Consent process

Identifying risk factors for preeclampsia in subsequent pregnancies in women with a history of preeclampsia

Preeclampsia (PE) is a prevalent hypertensive disorder of pregnancy that causes significant maternal-fetal mortality. It has been reported that 60% of maternal mortality can be prevented. Studies have demonstrated that nursing has the ability to influence the mortality rates by providing proactive and competent care. Part of proactive, competent care is to identify those women at the highest risk for PE. Multiple studies have identified a history of PE as a significant risk factor for PE in future gestations. We conducted a nested case control study with our Rule Out Pre-Eclampsia (ROPE) cohort to identify comorbidities most likely to lead to recurrent PE

Evaluating Quality Improvement to Improve HIV Reporting

The incorporation and evaluation of Quality Improvement into Georgia’s public health systems continues to be a focus of the Georgia Public Health Practice Based Research Network. This report describes the process, preliminary results and lessons learned from incorporating Quality Improvement into one of Georgia’s public health districts

The American Redstart (Setophaga ruticilla), insects, and foliage on Douglas Lake, Michigan.

BirdsThis study investigated whether there is a correlation between American Redstart (Setophaga ruticilla) distribution and foliage density, as well as a correlation between insect density and American Redstart distribution in a deciduous forest on Douglas Lake, Michigan. Birds were counted by observing and listening to American Redstarts as they sang early on four mornings at ten selected points in this forest. At the same ten points, understory foliage density was scored and sticky traps were set up to collect insects, spanning eight days. A linear regression and scatter plot with a best fit line calculated by SPSS revealed a significant positive correlation (R2 = 0.76) between the average number of American Redstarts at a given data collection point and its corresponding foliage density score, which supported the hypothesis that there would be more American Redstarts in areas with more foliage. The results concerning insects obtained from statistical analysis in SPSS did not support the hypothesis that there would be a positive correlation between arthropod density and American Redstart density. This information can be used in maintaining the habitat of the American Redstart and related species.http://deepblue.lib.umich.edu/bitstream/2027.42/64853/1/Benjamin_Davis_Donajkowski_Hall_Morris_Mosher_2009.pd

Appalachian Spring

A large mural that was created by 19 studio art and education students in 2009. Renowned Latina artist Judith Baca worked alongside the students.https://scholarworks.moreheadstate.edu/university_art_collection/1090/thumbnail.jp

A cross-sectional mixed methods study protocol to generate learning from patient safety incidents reported from general practice

Introduction Incident reports contain descriptions of errors and harms that occurred during clinical care delivery. Few observational studies have characterised incidents from general practice, and none of these have been from the England and Wales National Reporting and Learning System. This study aims to describe incidents reported from a general practice care setting. Methods and analysis A general practice patient safety incident classification will be developed to characterise patient safety incidents. A weighted-random sample of 12 500 incidents describing no harm, low harm and moderate harm of patients, and all incidents describing severe harm and death of patients will be classified. Insights from exploratory descriptive statistics and thematic analysis will be combined to identify priority areas for future interventions. Ethics and dissemination The need for ethical approval was waivered by the Aneurin Bevan University Health Board research risk review committee given the anonymised nature of data (ABHB R&D Ref number: SA/410/13). The authors will submit the results of the study to relevant journals and undertake national and international oral presentations to researchers, clinicians and policymakers

The X-Ray Crystal Structure of Escherichia coli Succinic Semialdehyde Dehydrogenase; Structural Insights into NADP+/Enzyme Interactions

In mammals succinic semialdehyde dehydrogenase (SSADH) plays an essential role in the metabolism of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) to succinic acid (SA). Deficiency of SSADH in humans results in elevated levels of GABA and gamma-Hydroxybutyric acid (GHB), which leads to psychomotor retardation, muscular hypotonia, non-progressive ataxia and seizures. In Escherichia coli, two genetically distinct forms of SSADHs had been described that are essential for preventing accumulation of toxic levels of succinic semialdehyde (SSA) in cells.Here we structurally characterise SSADH encoded by the E coli gabD gene by X-ray crystallographic studies and compare these data with the structure of human SSADH. In the E. coli SSADH structure, electron density for the complete NADP+ cofactor in the binding sites is clearly evident; these data in particular revealing how the nicotinamide ring of the cofactor is positioned in each active site.Our structural data suggest that a deletion of three amino acids in E. coli SSADH permits this enzyme to use NADP+, whereas in contrast the human enzyme utilises NAD+. Furthermore, the structure of E. coli SSADH gives additional insight into human mutations that result in disease