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Development And Applications Of Capillary Vibrating Sharp-Edge Spray Ionization Coupled With Atmospheric Pressure Chemical Ionization (Cvssi-Apci), Madison Elizabeth Pursell

Graduate Theses, Dissertations, and Problem Reports (ETD)

Mass spectrometry (MS) is a technique that is used in fields in which both complex and non-complex mixtures are analyzed. The state-of-the-art ion sources that are utilized for MS are being demonstrated to be functional for direct infusion of samples as well as for use with online separations. However, ion sources, such as electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) can encounter problems such as ion suppression and an inability to ionize based on size and/or polarity, respectively. Further these sources are bulky and require gas flow, heat sources, and additional high voltage power supplies for optimal performance. …

Positive Responses Of Solanum Lycopersicum Growth To Additions Of Gibberellin In The Presence Of Wounding Stress, Tatum Curtin

Undergraduate Scholarship

Herbivory is a consistent and prevalent stressor that threatens plants. Gibberellin, a naturally occurring plant growth hormone, has been known to increase plant growth when under a physical stress such as herbivory. To investigate, we set up a two-factorial design with 60 Solanum lycopersicum plants and subjected 15 plants each to four treatment groups: reference/control, 65 mg gibberellin, wounding, and 65 mg gibberellin+wounding. We found that S. lycopersicum root-to-shoot ratio decreases with the addition of gibberellin under mechanical wounding stress. Additionally, chlorophyll fluorescence increases under any mechanical wounding, with and without gibberellin present. Our experiment allows us to conclude that …

Advanced Techniques In Time Series Forecasting: From Deterministic Models To Deep Learning, Xue Bai

Graduate Theses, Dissertations, and Problem Reports (ETD)

This dissertation discusses three instances of temporal prediction, applied to population dynamics and deep learning.

In population modeling, dynamic processes are frequently represented by systems of differential equations, allowing for the analysis of various phenomena. The first application explores modeling cloned hematopoiesis in chronic myeloid leukemia (CML) via a nonlinear system of differential equations. By tracking the evolution of different cell compartments, including cycling and quiescent stem cells, progenitor cells, differentiated cells, and terminally differentiated cells, the model captures the transition from normal hematopoiesis to the chronic and accelerated-acute phases of CML. Three distinct non-zero steady states are identified, representing …

Conformational Investigation Of Biopolymers Through Molecular Dynamics (Md) Simulations And In-Droplet Hdx-Ms Utilizing Cvssi, Samira Hajian Foroushani

Graduate Theses, Dissertations, and Problem Reports (ETD)

Over the last two decades, mass spectrometry (MS) has received significant interest in biological study platforms, especially in the areas of proteomics and metabolomics. With the development of soft ionization methods like electrospray ionization (ESI), mass spectrometry has emerged as a potent method for directly characterizing proteins in their native state. The recent invention of capillary vibrating sharp-edge spray ionization has opened new avenues for the study of biopolymer native structure including the usage of droplet charge tuning and in-droplet hydrogen-deuterium exchange (HDX) labeling. With this new technique comes the opportunity to study biopolymer ion structure preservation with molecular dynamics …

Syntheses Of Indoles And Natural Indole Alkaloids From 2-Nitrostyrene Derivatives, S M Ashikur Rahman

Graduate Theses, Dissertations, and Problem Reports (ETD)

A short and flexible synthesis of 2,3-disubstituted indoles employing a Barluenga-type palladium-catalyzed cross-coupling of p-tosylhydrazones with 2-nitroarylhalides followed by a palladium-catalyzed, carbon monoxide-mediated reductive cyclization was developed. Significantly better overall yields were obtained performing the two-step reaction sequence in a one-pot reaction. Secondly, in order to facilitate biological studies of ergotryptamine, a recently isolated ergot alkaloid, it was selected as a target for synthesis. In an attempt to synthesize ergotryptamine, a palladium catalyzed Kosugi-Migita-Stille cross coupling, a Mizoroki-Heck reaction, and a carbon monoxide-mediated palladium-catalyzed reductive cyclization were utilized as the key steps. Using a similar synthetic approach, synthesis of …

Biocompatible And Multifunctional Trityl Spin Probes For Electron Paramagnetic Resonance Spectroscopy, Teresa D. Gluth

Graduate Theses, Dissertations, and Problem Reports (ETD)

The primary objective of my thesis was to develop and utilize a biocompatible multifunctional trityl spin probe for concurrent measurement of pO₂, pH_e, and [P_i] in vivo by electron paramagnetic resonance (EPR) spectroscopy (Chapter 2). My first goal was to synthesize the proposed probe we are terming HOPE71. Secondly, HOPE71 was characterized by X-band and L-band EPR spectroscopy. Next, the biocompatibility of HOPE71 was assessed through an albumin binding test, cytotoxicity assays, and in vivo intravenous tolerance. Then, the use of HOPE71 to measure the target parameters was demonstrated in a breast cancer …

Harnessing Acoustic Streaming For Bioanalysis In Microfluidics, Xiaojun Li

Graduate Theses, Dissertations, and Problem Reports (ETD)

In the past few decades, microfluidic technology has been developed rapidly in both fabrication methods and multifunctionality integrations, making it a powerful tool for a wider variety of biological applications. The device fabrication method has expanded from conventional polymer-based micro devices to 3D-printed micro devices, and allows multiple functions such as electric, optic, magnetic and acoustic to be integrated on a single platform. This dissertation focuses on method development of acoustic streaming-based microfluidics in bioanalysis such as immunoassay, enzyme kinetics and DNA fragmentation analysis. In this dissertation, a micromixer was developed based on acoustic streaming generated from sharp-edge structure vibration …

Assessment Of Heat Stress For Outdoor Work Conditions In Saudi Arabia, Jamal Alanazi

Graduate Theses, Dissertations, and Problem Reports (ETD)

Outdoor workers have an increased risk of heat stress in Saudi Arabia since it is one of the hottest places in the Middle East. Recently, the government decided to limit outdoor work hours during the months of June, July, and August every year, and banned working under the direct sunlight from 12:00 to 03:00 p.m., although outdoor workers in the petroleum, natural gas, or emergency maintenance work industries are exempt from this prohibition. Traditionally, the efforts by safety and health professionals to mitigate work-related heat injury has been directed toward the assessment of environmental heat stress (e.g., wet-bulb globe temperature), …

The Role Of Genotype In Saccharomyces Cerevisiae Resistance To 4-Methylcyclohexanemethanol, Michael C. Ayers

Graduate Theses, Dissertations, and Problem Reports (ETD)

The chemical 4-methylcyclohexanemethanol (MCHM) spilled in the Elk River near Charleston, WV in January 2014, impacting the drinking water of 300,000 residents. Initial studies on MCHM following the spill attempted to characterize its toxicological effects in animals, but largely ignored characterization of its effects on cellular pathways and metabolism. In this study, I used this chemical as a novel stressor for the model eukaryote S. cerevisiae to explore the relationship of genotype and stress phenotypes. Initial exploration characterized the stress phenotype of MCHM in yeast through growth assays, cell cycle analysis, and transcriptomic data. MCHM exposure caused cells to arrest …

Protein/Peptide Characterization Using Mass Spectrometry And Molecular Dynamics Simulations, Ahmad Kiani Karanji

Graduate Theses, Dissertations, and Problem Reports (ETD)

Mass spectrometry (MS) based-techniques and molecular dynamics (MD) simulations have been used to characterize protein/peptide structure as well as their interactions with lipid vesicles and detergents. Chapter 1 introduces an introduction to the concepts and tools that were used in this work. In Chapter 2, the dominant gas-phase conformer of [M+3H]3+ ions of the model peptide Acetyl-PSSSSKSSSSKSSSSKSSSSK are examined with ion mobility spectrometry (IMS), gas-phase hydrogen deuterium exchange (HDX), and mass spectrometry (MS) techniques. This section furthers the development of a protein structural prediction tool by providing information about gas-phase ion conformers of two model peptides having different solution conformational …

The Stem Cell Phenotype Of Aggressive Breast Cancer Cells, Naira V. Margaryan, Hannah Hazard-Jenkins, Mohamad A. Salkeni, Matthew B. Smolkin, James A. Coad, Sijin Wen, Elizabeth A. Seftor, Richard E B Seftor, Mary J C Hendrix

Faculty & Staff Scholarship

Aggressive cancer cells are characterized by their capacity to proliferate indefinitely and to propagate a heterogeneous tumor comprised of subpopulations with varying degrees of metastatic propensity and drug resistance properties. Particularly daunting is the challenge we face in the field of oncology of effectively targeting heterogeneous tumor cells expressing a variety of markers, especially those associated with a stem cell phenotype. This dilemma is especially relevant in breast cancer, where therapy is based on traditional classification schemes, including histological criteria, differentiation status, and classical receptor markers. However, not all patients respond in a similar manner to standard-of-care therapy, thereby necessitating …

How To Search For Life In Martian Chemical Sediments And Their Fluid And Solid Inclusions Using Petrographic And Spectroscopic Methods, Kathleen C. Benison

Faculty & Staff Scholarship

Abundant resources and efforts have been employed in the search for life on Mars. Satellites, landers, and rovers have tested atmospheric gases, general sediment and rock compositions, and images of Mars surface in an effort to detect biosignatures left by any possible modern or ancient life. Chloride and sulfate minerals suggestive of past acid saline lakes have been found on Mars. In terrestrial acid brine environments, these minerals trap microorganisms and organic compounds and preserve them within fluid inclusions and as solid inclusions for long geologic time periods. Some cells remain viable, especially in the isolated, microscopic aqueous environments of …

Lpa Signaling Is Regulated Through The Primary Cilium: A Novel Target In Glioblastoma, Yuriy V. Loskutov, Caryn L. Griffin, Kristina M. Marinak, Andrey Bobko, Naira V. Margaryan, Werner J. Geldenhuys, Jann N. Sarkaria, Elena N. Pugacheva

Clinical and Translational Science Institute

The primary cilium is a ubiquitous organelle presented on most human cells. It is a crucial signaling hub for multiple pathways including growth factor and G-protein coupled receptors. Loss of primary cilia, observed in various cancers, has been shown to affect cell proliferation. Primary cilia formation is drastically decreased in glioblastoma (GBM), however, the role of cilia in normal astrocyte or glioblastoma proliferation has not been explored. Here we report that loss of primary cilia in human astrocytes stimulates growth rate in a lysophosphatidic acid (LPA)-dependent manner. We show that lysophosphatidic acid receptor 1 (LPAR1) is accumulated in primary cilia. …

Design And Synthesis Of Small Molecule Agonists Of Epha2 Receptor, Aaron Petty, Nethrie Idippily, Viharika Bobba, Werner J. Geldenhuys, Bo Zhong, Bin Su, Bingcheng Wang

Clinical and Translational Science Institute

Ligand-independent activation of EphA2 receptor kinase promotes cancer metastasis and invasion. Activating EphA2 receptor tyrosine kinase with small molecule agonist is a novel strategy to treat EphA2 overexpressing cancer. In this study, we performed a lead optimization of a small molecule Doxazosin that was identified as an EphA2 receptor agonist. 33 new analogs were developed and evaluated; a structure−activity relationship was summarized based on the EphA2 activation of these derivatives. Two new derivative compounds 24 and 27 showed much improved activity compared to Doxazosin. Compound 24 possesses a bulky amide moiety, and compound 27 has a dimeric structure that is …

Allicin- Inspired Pyridyl Disulfides As Antimicrobial Agents For Multidrug-Resistant Staphylococcus Aureus, Jordan G. Sheppard, Jeremy P. Mcaleer, Pushkar Saralkar, Werner J. Geldenhuys, Timothy E. Long

Clinical and Translational Science Institute

A chemical library comprised of nineteen synthesized pyridyl disulfides that emulate the chemical reactivity of allicin (garlic) was evaluated for antimicrobial activity against a panel of pathogenic bacteria. Gram-positive species including vancomycin-intermediate and vancomycin-resistant Staphylococcus aureus (VISA, VRSA) demonstrated the highest level of susceptibility toward analogs with S-alkyl chains of 7–9 carbons in length. Further biological studies revealed that the disulfides display synergy with vancomycin against VRSA, cause dispersal of S. aureus biofilms, exhibit low cytotoxicity, and decelerate S. aureus metabolism. In final analysis, pyridyl disulfides represent a novel class of mechanism-based antibacterial agents that have a potential application as …

Graphene Nanoplatelets-Sericin Surface-Modified Gum Alloy For Improved Biological Response, Valentina Mitran, Valentina Dinca, Raluca Ion, Vasile D. Cojocaru, Patricia Neacsu, Cerasela Zoica Dinu, Laurentiu Rusen, Simona Brajnicov, Anca Bonciu, Maria Dinescu, Doina Raducanu, Ioan Dan

Faculty & Staff Scholarship

In this study a “Gum Metal” titanium-based alloy, Ti-31.7Nb-6.21Zr-1.4Fe-0.16O, was synthesized by melting and characterized in order to evaluate its potential for biomedical applications. The results showed that the newly developed alloy presents a very high strength, high plasticity and a low Young's modulus relative to titanium alloys currently used in medicine. For further bone implant applications, the newly synthesized alloy was surface modified with graphene nanoplatelets (GNP), sericin (SS) and graphene nanoplatelets/sericine (GNP–SS) composite films via Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique. The characterization of each specimen was monitored by scanning electron microscopy (SEM), atomic force microscopy (AFM), …

Syntheses Of Azepinoindole Alkaloids Cimitrypazepine, Fargesine And Hyrtioreticulins C & D, Ganesh Ghimire

Graduate Theses, Dissertations, and Problem Reports (ETD)

The Watanabe-Cenini-Söderberg reductive N-heterocyclization has emerged as a powerful tool for the synthesis of a variety of functionalized indoles. Söderberg’s elaboration of this methodology has been utilized as a late-stage cyclization protocol for the synthesis of azepinoindole alkaloids, a class of natural products characterized by an azepane skeleton fused with an indole ring. Short syntheses of the naturally occurring azepino[5,4,3-cd]indole alkaloids, cimitrypazepine, fargesine, and the diastereomeric hyrtioreticulins C & D have been completed starting from commercially available tetra-substituted benzene derivatives. The key azepinoindole core in each case was assembled via an intramolecular Mizoroki-Heck reaction followed by a …

In Vitro Recapitulation Of Functional Microvessels For The Study Of Endothelial Shear Response, Nitric Oxide And [Ca2+]I, Xiang Li, Sulei Xu, Pingnian He, Yuxin Liu

Faculty & Staff Scholarship

Microfluidic technologies enable in vitro studies to closely simulate in vivo microvessel environment with complexity. Such method overcomes certain constrains of the statically cultured endothelial monolayers and enables the cells grow under physiological range of shear flow with geometry similar to microvessels in vivo. However, there are still existing knowledge gaps and lack of convincing evidence to demonstrate and quantify key biological features of the microfluidic microvessels. In this paper, using advanced micromanufacturing and microfluidic technologies, we presented an engineered microvessel model that mimicked the dimensions and network structures of in vivo microvessels with a long-term and continuous perfusion capability, …

But Who Will Get Billy? The Effect Of Child Custody Laws On Marriage, Elaina Rose, Crystal (Ho Po) Wong

Economics Faculty Working Papers Series

Under the tender years doctrine in effect until the 1970’s, custody was virtually always awarded to the mother upon divorce. Gender-neutral custody laws introduced beginning in the 1970’s provided married fathers, in principle, equal rights to custody. Subsequent marriage-neutral laws extended the rights to unmarried fathers. We develop a theoretical model of the effect of custody regime on marriage and test the model’s predictions using a unique data set that merges custody law data with data from the Current Population Survey and Vital Statistics. We find that, under marriage non-neutrality, the introduction of gender-neutral laws reduced the hazard into marriage …

Multiple Mineralocorticoid Response Elements Localized In Different Introns Regulate Intermediate Conductance K+ (Kcnn4) Channel Expression In The Rat Distal Colon, Bryan O'Hara, Diego A. Rosa, Vazhaikkurichi M. Rajendran

Faculty & Staff Scholarship

An elevated plasma aldosterone and an increased expression of the intermediate conductance K+ (IK/Kcnn4) channels are linked in colon. This observation suggests that the expression of Kcnn4 gene is controlled through the action of aldosterone on its cognate receptor (i.e., mineralocorticoid receptor; MR). In order to establish this, we performed chromatin immunoprecipitation (ChIP) assay to identify the MR response elements (MREs) in a region that spanned 20 kb upstream and 10 kb downstream of the presumed transcription start site (TSS) using chromatin from the colonic epithelial cells of normal and aldosterone-treated rats. MREs were immunoprecipitated in an approximately 5 kb …

Living Microbe Repository, Best Practices, Regulatory Compliance, Reference Material Collection, Shaolei Zang, Na Liu, Hongchun Wang, David N. Wald, Na Shao, Jingru Zhang, Daoxin Ma, Chunyan Ji, William Tse

Faculty & Staff Scholarship

Background

We previously demonstrated that 6-benzylthioinosine (6-BT) could induce the differentiation of a subset of acute myeloid leukemia (AML) cell lines and primary AML cells regardless of their cytogenetics. In this study we investigated whether Wnt signaling pathways played roles in 6-BT-induced differentiation of AML cells.

Methods

We induced differentiation of HL-60 leukemic cells and primary AML cells in vitro using 6-BT. Real-time PCR (qPCR), western blot, and luciferase assays were used to examine the molecules’ expression and biological activity in canonical and noncanonical Wnt signaling pathways. AML cell differentiation was measured by the Nitroblue tetrozolium (NBT) reduction assay.

Results …

Investigating The Role Of Mechanical Properties In Biological Systems, Nicole Shamitko-Klingensmith

Graduate Theses, Dissertations, and Problem Reports (ETD)

There is considerable interest in measuring, with nanoscale spatial resolution, the physical and material properties of biological membranes and whole cells because of their role in the physiology of living systems. The atomic force microscope (AFM) has proven to be particularly well-suited for biological studies because samples can be maintained in near physiological conditions to preserve sample integrity (such as in a buffer solution at a physiologically relevant pH or temperature), and the imaging process is relatively nondestructive. In this work, the mechanical properties of supported lipid bilayers and neurons were examined using AFM-based techniques. Specifically, tapping mode AFM-based techniques …

Notch1 Is Required For Hypoxia-Induced Proliferation, Invasion And Chemoresistance Of T-Cell Acute Lymphoblastic Leukemia Cells, Jie Zou, Peng Li, Fei Lu, Na Liu, Jianjian Dai, Jingjing Ye, Xun Qu, Xiulian Sun, Daoxin Ma, Jino Park, Chunyan Ji

Faculty & Staff Scholarship

Background

Notch1 is a potent regulator known to play an oncogenic role in many malignancies including T-cell acute lymphoblastic leukemia (T-ALL). Tumor hypoxia and increased hypoxia-inducible factor-1α (HIF-1α) activity can act as major stimuli for tumor aggressiveness and progression. Although hypoxia-mediated activation of the Notch1 pathway plays an important role in tumor cell survival and invasiveness, the interaction between HIF-1α and Notch1 has not yet been identified in T-ALL. This study was designed to investigate whether hypoxia activates Notch1 signalling through HIF-1α stabilization and to determine the contribution of hypoxia and HIF-1α to proliferation, invasion and chemoresistance in T-ALL.

Methods …

Evaluation Of Gene Association Methods For Coexpression Network Construction And Biological Knowledge Discovery, Sapna Kumari, Jeff Nie, Huann-Sheng Chen, Hao Ma, Ron Stewart, Xiang Li, Meng-Zhu Lu, William M. Taylor, Hairong Wei

Faculty & Staff Scholarship

Background

Constructing coexpression networks and performing network analysis using large-scale gene expression data sets is an effective way to uncover new biological knowledge; however, the methods used for gene association in constructing these coexpression networks have not been thoroughly evaluated. Since different methods lead to structurally different coexpression networks and provide different information, selecting the optimal gene association method is critical.

Methods and Results

In this study, we compared eight gene association methods – Spearman rank correlation, Weighted Rank Correlation, Kendall, Hoeffding's D measure, Theil-Sen, Rank Theil-Sen, Distance Covariance, and Pearson – and focused on their true knowledge discovery rates …

Microfluidic Impedance-Metric Biosensor For Monitoring Carbon Nanotube-Induced Epithelial Permeability, Zhouchun Huang

Graduate Theses, Dissertations, and Problem Reports (ETD)

Carbon nanomaterials, including carbon nanotubes (CNTs) gain increasing interest in commercial and biomedical applications. Adverse health effects of engineered CNTs may result from either direct exposure during production, inadvertent exposure during use of by-products, and/or release of these materials into the environment. Recent studies have shown that CNTs can induce cytotoxicity, apoptosis, DNA damage, and tumor formation in animals, suggesting their potential toxicity in humans. To address the current needs in developing in vitro cell assays for assessment of the adverse health effects of CNTs in lung respiratory, we have developed a microfluidic impedance-metric biosensor for investigation of multi-walled CNTs …

Surfaces Modulate Β-Amyloid Peptide Aggregation Associated With Alzheimer's Disease., Elizabeth Anne Yates

Graduate Theses, Dissertations, and Problem Reports (ETD)

A hallmark of Alzheimer's disease, a late onset neurodegenerative disease, is the presence of neuritic amyloid plaques deposited within the brain composed of β-amyloid (Aβ) peptide aggregates. Aβ can aggregate into a variety of polymorphic aggregate structures under different chemical environments, specifically affected by the presence of differing surfaces. There are several point mutations clustered around the central hydrophobic core of Aβ (E22G Arctic mutation, E22K Italian mutation, D23N Iowa mutation, and A21G Flemish mutation). These mutations are associated with hereditary diseases ranging from almost pure cerebral amyloid angiopathy to typical Alzheimer's disease pathology with both plaques and tangles. To …

Pharmacological Characterization Of An Optimized Sigma Receptor Ligand, Michael Seminerio

Graduate Theses, Dissertations, and Problem Reports (ETD)

Methamphetamine is an addictive psychostimulant drug with an increasing prevalence of abuse worldwide. Recent evidence has linked methamphetamine exposure to a wide array of neurological complications some of which lead to neurodegenerative changes in the brain and hyperthermia. Currently, there are no FDA approved pharmacologic agents to treat the effects of methamphetamine. Recent evidence has suggested methamphetamine may, in part, produce some of its effects through sigma receptors making them a potential target for pharmacological intervention. While our lab has shown a number of selective sigma receptor ligands can provide neuroprotection against the neurotoxic and behavioral effects of methamphetamine, limitations …

The Relationship Between Temperament And Facets Of Creativity, Clark Daniel Pinson

Graduate Theses, Dissertations, and Problem Reports (ETD)

The purported association between mental illness and creativity has been most heavily researched in individuals diagnosed with a bipolar disorder. The bipolar disorders have been conceptualized as a spectrum of disorders that range from problematic and labile affect to different types of temperament. Temperament has classically been defined as elements of personality that are heritable, stable, based on emotion, and uninfluenced by socio-cultural learning. Contemporary literature suggested that at least some aspects of creativity may be enhanced by an affective range that has been best characterized by the milder part of the bipolar spectrum of disorders. The primary inquiry of …

Assessment Of Pulmonary Fibrogenic Potential Of Multi Walled Carbon Nanotubes In Vitro., Anurag Mishra

Graduate Theses, Dissertations, and Problem Reports (ETD)

Multi-walled carbon nanotubes (MWCNT) have many applications due to their unique mechanical and electrical properties. However, these fiber shaped materials have been shown to possess unusual fibrogenic activity in vivo and are currently the focus of intense toxicological investigations. To date very few detailed mechanistic studies on the fibrogenic activity of CNT have been reported. The aim of this series of studies is to determine fibrogenic potential of well dispersed MWCNT in human lung cell culture models and develop a novel platform for understanding the cellular mechanisms of MWCNT-induced lung fibrosis. Dispersion of large agglomerates of MWCNT in suspension to …

Computational Hybrid Systems For Identifying Prognostic Gene Markers Of Lung Cancer, Ying-Wooi Wan

Graduate Theses, Dissertations, and Problem Reports (ETD)

Lung cancer is the most fatal cancer around the world. Current lung cancer prognosis and treatment is based on tumor stage population statistics and could not reliably assess the risk for developing recurrence in individual patients. Biomarkers enable treatment options to be tailored to individual patients based on their tumor molecular characteristics. To date, there is no clinically applied molecular prognostic model for lung cancer. Statistics and feature selection methods identify gene candidates by ranking the association between gene expression and disease outcome, but do not account for the interactions among genes. Computational network methods could model interactions, but have …