Physical Sciences and Mathematics Commons^™

Targeting Heat Shock 27 Kda Protein Induces Androgen Receptor Degradation, Yaxin Li

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Glioblastoma (GBM) is the most common and aggressive brain tumor, with very poor prognosis. Androgen receptor (AR) plays a significant role in the progression of GBM, and anti-androgen agents have the potential to be used for the treatment of GBM. However, AR mutation commonly happens in GBM, which makes the anti-androgen agents less effective. Heat shock 27 kDa protein (HSP27) is a well-documented chaperone protein to stabilize AR. Inhibition of HSP27 results in AR degradation regardless the mutation status of AR, which makes HSP27 a good target to abolish AR in GBM. Identified compound I ((N-(3-((2,5-dimethoxybenzyl)oxy)-4-(methylsulfonamido) phenyl)-4-methoxybenzamide) inhibits GBM cell …

Biochemical Characterization Of Induced Pluripotent Stem Cell-Derived Cardiomyocytes As A Model Of Barth Syndrome, Alisha J. House

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Barth Syndrome (BTHS) is an X-linked inborn error of metabolism (IEM) which manifests as a multi-systemic disease. One of the primary symptoms is dilated cardiomyopathy, and alongside the cardiovascular disease that arises, patients often experience metabolic abnormalities such as 3-methylglutaconic aciduria, growth retardation, and neutropenia. There has been a need for the development of a suitable in vitro modeling system which will accurately recapitulate the biochemical and physical nature of BTHS. The purpose of this project has been to develop a model for studying the biochemical pathogenesis of Barth Syndrome using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). To achieve …

Synthesis And Functionality Study Of Novel Biomimetic N-Glycan Polymers, Ka Keung Chan

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Cell surface carbohydrates, existing as glycoproteins, glycolipids, and proteoglycans, often serve as receptors to relay critical interactions and trigger crucial signaling events in many biological processes. Glycan recognition is deeply involved in immune surveillance, response and can provide abundant opportunities to discover the molecular mechanism and potential therapeutic or diagnostic approaches for various diseases. Many useful functionalities can be achieved by developing synthetic glycans to mimic natural carbohydrates. Over the recent decade, glycopolymers, polymers with pendant carbohydrates, have continued to prove usefulness in various applications across different scientific disciplines. For example, glycopolymers can be used as biomimetic glycoligands to study …

Synthesis, Characterization, And Catalytic And Biological Activities Of A Mixed-Ligand Cobalt(Ii) Bipyridyl/Diphenylazodioxide Complex, Kylin A. Emhoff

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Transition metal complexes have immense importance in the pharmaceutical industry. These types of complexes can be useful catalysts in the synthesis of medicinal compounds and can act as anticancer drugs. In these pharmaceutical applications, 1st-row transition metal-containing complexes offer certain advantages compared to their 2nd and 3rd-row transition metal counterparts. Our motivation was to investigate pharmaceutical applications of transition metal complexes containing both a 1st-row transition metal and unusual ligands to expand the knowledge of a class of complexes that could potentially be beneficial in the pharmaceutical industry. A class of rare ligands that piqued our interest was that of …

Determination Of Dynorphins And Tnf- A By Lc-Ms/Ms In Biological Samples: Applicable To Studying Inflammatory Mechanisms, Karthik Chandu

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Dynorphins are endogenous opioid peptides that have been implicated as initiators ofimmune and inflammatory response through upregulation ofinflammatory cytokine and chemokine production, as well having a role in glutamate-induced neuro-inflammation and neurotoxicity. Being extremely potent peptides, the physiologic concentrations of dynorphins are very low ranging from 0.16 pg/mL during the absence of a stimulus to 23.5 pg/mL when stimulated in disease condition. Previously published HPLC-mass spectrometry techniques have insufficient detection capabilities for quantification and detection ofdynorphins. As a result, immunoassay quantification has been the most utilized technique for analysis of dynorphins in physiologic samples. Although being sensitive, immunoassays have some …

Expression And Cell Surface Re-Engineering Of Thrombomodulin On Macrophages, Mallorie L. Boron

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Thrombomodulin (TM) is a transmembrane glycoprotein that is primarily expressed on the surface of endothelial cells, where it serves as a receptor of thrombin for protein C activation, which regulates coagulation and inflammation. Research has revealed that TM is also expressed in immune cells, including monocytes and macrophages, however, its function is unclear. In this dissertation study, the TM expression in THP-1 monocytes upon their differentiation to macrophages was profiled. Due to the beneficial roles of TM, the protein has caught the attention to be used as a therapeutic. A recombinant form of TM can be expressed that contains EGF-like …

Coordination Chemistry Of N,N’- Azodoxides, Lakshmi Balaraman

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The coordination chemistry of nitric oxide (NO) with transition metals has been extensively investigated, but that of organic NO derivatives as ligands less so. Some nitrosoalkanes RNO and nitrosoarenes ArNO are in equilibrium with dimeric species known as N,N'–azodioxides, R(O)NN(O)R or Ar(O)NN(O)Ar. The cis diastereomers of azodioxides have been proposed to act as bidentate chelating ligands for both main-group and transition metals, but only three azodioxide complexes, a dicationic iron (II) complex, a neutral scandium (III) complex, and a dicationic calcium (II) complex, have thus far been structurally characterized before our work in this area. The delocalized π system of …

Reactivity Of Molybdenum And Tungsten Sulfido Complexes With First-Series Transition Metals, Khalil Mudarmah

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Ammonium tetrathiomolybdate (ATTM) is an important compound in bioinorganic chemistry, with medicinal applications. Metal complexes of sulfur-containing ligands have been widely used as building block in biology and inorganic chemistry. This project aims to increase the number of Mo-S and W-S complexes that may contribute to fields such as bioinorganic and medicinal chemistry, by the preparation of heterometallic complexes containing both Mo or S and a metal of the first transition series. The homobimetallic complex anion salts (Et4N)2[Mo2(S)2(μ-S)2(edt)2] (1) and (Et4N)2[W2(S)2(μ-S)2(edt)2] (2) have been prepared from ATTM and ammonium tetrathiotungstate (ATTW) in order to start our work. The overnight treatment …

Liquid Chromatography - Mass Spectrometric Analysis Of Clinicallyand Pharmacologically Relevant Molecules, Raghavi Kakarla

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Liquid Chromatography-Mass Spectrometry is an advanced analytical technique that offers high sensitivity and specificity and has been increasingly used for analysis of a wide variety of compounds including clinically and pharmacologically relevant molecules. In this dissertation we describe qualitative and quantitative liquid chromatographic mass spectrometric methods to analyze both small molecules and larger macromolecules that provide useful insights into diagnosis and management of several diseases. An LCMS(/MS) analytical method includes extraction of analytes of interest from the matrix followed by liquid chromatographic separation and mass spectrometric detection. Chapter I describes pre-analytical workflows and sample pretreatment techniques and theories underlying LC-MS …

Development Of An Spr Method To Monitor The Chemical Interaction Of Dna With A Small Reactive Molecule: Peroxynitrite As An Example, Divya Sharma

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Peroxynitrite is an anion with the formula ONOO-. It is an unstable structural isomer of nitrate, NO3-. Although its conjugate acid (ONOOH) is highly reactive, it is quite relatively stable in basic solutions. Generation of peroxynitrite in-vivo occurs through the diffusion-controlled reaction between superoxide anion and nitric oxide free radical. It is a strong oxidizing and nitrating agent, and its formation has been correlated with many pathological conditions. Because of its oxidizing properties, peroxynitrite can damage a wide array of molecules in cells, including DNA and proteins. Reactions of peroxynitrite with DNA, proteins, and lipids, trigger a wide array of …

In-Quest Of Biomarkers For Alzheimer’S Disease And Pharmacokinetic Profile Of Anticancer Agents Using Lc-Ms In Human Plasma, Chandana Mannem

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In this dissertation work, a semi-quantitative assay using a lipidomics approach and an absolute quantitative assay using liquid chromatography-mass spectrometry techniques were developed (LC-MS). In the lipidomics approach, ultra-high pressure liquid chromatography in tandem with quadrupole time of flight (UHPLC-QTOF) mass spectrometry was used to profile, compare and quantitate the human plasma lipids in Alzheimer’s disease subjects (AD) and normal cognitive controls (NCCs). The purpose of this study is to identify potential plasma lipid markers of AD and to explore the relationships between AD and lipid pathways in humans by using bioinformatic tools. The plasma samples of study subjects were …

Development Of An Spr Method To Monitor The Chemical Interaction Of Dna With A Small Reactive Molecule: Peroxynitrite As An Example, Divya Sharma

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Peroxynitrite is an anion with the formula ONOO-. It is an unstable structural isomer of nitrate, NO3-. Although its conjugate acid (ONOOH) is highly reactive, it is quite relatively stable in basic solutions. Generation of peroxynitrite in-vivo occurs through the diffusion-controlled reaction between superoxide anion and nitric oxide free radical. It is a strong oxidizing and nitrating agent, and its formation has been correlated with many pathological conditions. Because of its oxidizing properties, peroxynitrite can damage a wide array of molecules in cells, including DNA and proteins. Reactions of peroxynitrite with DNA, proteins, and lipids, trigger a wide array of …

Development Of Bioanalytical Methods For Quantitative Measurement Of Anticancer Agents, Sandeep R. Kunati

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The ever-growing need to develop new anticancer agents due to increased incidence of cancer and its recurrence has led to a broad range of investigational efforts in anticancer research. Drug discovery and development processes involve evaluations of pharmacokinetic and pharmacodynamic parameters of potential drug candidates in order to assess their safety, toxicity, and efficacy profiles. By providing sensitivity, assay precision, reliability, and high-throughput, mass spectrometry has become an indispensable tool in quantitative and qualitative analysis, from early-stage drug discovery and development processes to routine laboratory analysis. Development of bioanalytical methods requires knowledge of separation science with instrumentation, data interpretation and …

Biomimetic Macromolecules For Macrophage Targeting And Modulation, Joshua Whited

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Carbohydrate recognition has come to the forefront of biological aiming to uncover the mechanisms of physiological and pathological processes. Cell surface glycans are involved in processes including cellular adhesion, cell signaling, and immune response. A new approach for profiling cell surface glycans has great potential for a wide range of biomedical applications. Lectins have been conventionally used to determine the structure and function of glycoproteins, however, their numbers are still restricted compared to the number of glycan structures. Boronic acid has proven a remarkable small molecule capable of binding diols in aqueous solution. This interaction indicates boronic acid derived molecules …

Synthesis And Biological Evaluation Of Small Molecular Drug Candidates For The Treatment Of Her2 Overexpressed Breast Cancer, Anran Zhao

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In the US breast cancer is the most common cancer after skin cancer. Currently in the US the average risk of a woman to develop breast cancer in her life is roughly 12%. About 25-30% of breast cancer patients have HER2 overexpressed tumor, and the growth of tumor cell is depend on HER2 pathway. It has been well-documented that patients with over-expressed HER2 are associated with increased disease recurrence, worse prognosis and lower survival. Currently there are two type of HER2 targeting drug. The first group is HER2 monoclonal antibody drugs such as trastuzumab approved by FDA in 1998; the …

Quantitative Analysis Of Bleomycin In Rat Plasma By Lc-Ms/Ms, Huawen Li

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Bleomycin is the most commonly used compound in its group of antineoplastic drugs. It works on tumor cells by single and double stranded DNA cleavage after its activation, in which it blocks tumor cells’ DNA replication or transcription activities to inhibit tumor cells’ growth. Bleomycin sulfate (Blenoxane) is the most popular preparation used in clinical research, and contains Bleomycin fractions of A2 and B2, which causes difficulties in quantitative analysis. This work uses the metal chelating property of Bleomycin as an advantage to simplify and improve sensitivity of existing quantitative methods. Copper was spiked in excess to plasma samples, followed …

Computational Investigation Of Protein Assemblies, Sm Bargeen Alam Turzo

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Selective nitrosylation of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) at Cys-247 affects gene regulation through the interferon-gamma (IFN- γ) activated inhibitor of translation (GAIT) complex. Oxidized low-density lipoprotein (LDL_ox) and INF-γ induce assembly of the nitrosylase complex composed of inducible nitric oxide synthase (iNOS), S100A8, and S100A9 proteins. Crystal structure of the complex of GAPDH and S100A8A9 is not known, structural prediction method were employed by protein-protein docking and binding energy calculation with PatchDock and FIREDock respectively. Candidate models were selected, based on a weight factor calculated, from the computational method developed from the "artificial protease" cleavage mapping Fe(III) (s)-1-(p- …

Oxidation Of Ferrocene Derivatives With Dibenzoyl Peroxide And Meta-Chloroperoxybenzoic Acid, Jos M. Halstead

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The chemical oxidation of ferrocene and related derivatives (RFc) via organic peroxides solvated in acetonitrile was studied spectrophotometrically by varying concentration and temperature to determine kinetics and activation parameters. The reaction rate of ferrocene with dibenzoyl peroxide depends strongly on whether electron withdrawing or donating substituents are present. Products were analyzed and the effect of different solvents on reactivity were studied. The rate law was first order in both oxidant and reductant, and steric and solvent effects are consistent with outer sphere electron transfer (ET) as the rate-controlling step. B3YLP calculations were conducted to determine reorganization energies using Marcus theory …

Model Chemistry Study Of Choline And Urea Based Deep Eutectic Solvents, Libby Nicole Kellat

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Gaussian and GaussView software were utilized to characterize interactions between choline salts and urea, which form a deep eutectic solvent (DES). The initial system studied was choline chloride and urea, at a 1:2 molar ratio, which is also known as reline. Subsequent systems, substituting the chloride anion with other anions (fluoride, bromide, and hydroxide), were studied to show that the system with greater calculated strength of interaction will have more non-ideal physical properties, such as melting point (found in literature). Observations regarding structure related to counterion electron density and hydrogen bonding were made throughout the studies.

Il-17 Drives Copper Uptake And Activation Of Growth Pathways In Colorectal Cancer Cells In A Steap4-Dependent Manner, Evan Martin

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Colorectal cancer is a disease characterized by abnormal, invasive cell growth beginning in the colon or rectum. The third most common type of cancer worldwide, approximately one million new cases of the disease are diagnosed across the globe annually, resulting in an estimated 700,000+ deaths. One major risk factor associated with development of colorectal cancer is the presence of chronic inflammation in the large intestine, also known as colitis. Inflammation is a complex immune response against harmful stimuli, characterized by symptoms including heat, redness, swelling and pain. One important molecular mediator of this process is interleukin 17 (IL-17), a pro-inflammatory …

Il-17 Drives Copper Uptake And Activation Of Growth Pathways In Colorectal Cancer Cells In A Steap4-Dependent Manner, Evan Martin

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Colorectal cancer is a disease characterized by abnormal, invasive cell growth beginning in the colon or rectum. The third most common type of cancer worldwide, approximately one million new cases of the disease are diagnosed across the globe annually, resulting in an estimated 700,000+ deaths. One major risk factor associated with development of colorectal cancer is the presence of chronic inflammation in the large intestine, also known as colitis. Inflammation is a complex immune response against harmful stimuli, characterized by symptoms including heat, redness, swelling and pain. One important molecular mediator of this process is interleukin 17 (IL-17), a pro-inflammatory …

Using Nucleotide Analogs As Biochemical Probes To Evaluate The Mechanisms Involved In Translesion Replication By A High Fidelity Dna Polymerase, Anvesh Dasari

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Translesion DNA synthesis (TLS) allows DNA polymerases to incorporate nucleotides opposite and beyond damaged DNA. This activity is an important risk factor for the initiation and progression of genetic diseases including cancer. My study evaluates the ability of a high-fidelity DNA polymerase to perform TLS with 8-oxo-guanine, a pro-mutagenic DNA lesion formed by reactive oxygen species. Using modified purine and non-natural indole analogs as biochemical probes, I have evaluated the influence of desolvation, hydrogen bonding interactions, and shape complementarity towards nucleotide binding and incorporation opposite the miscoding lesion 8-oxo-guanine by the high fidelity gp43exo- DNA polymerase. In Chapter II, I …

Modified Electrodes With Grafted Dna And Oligonucleotides For Detection And Quantification Of Peroxynitrate, Heba Azmy Salim

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Peroxynitrite (ONOO-) is a strong oxidizing and nitrating agent, and its formation has been correlated with many pathological conditions. It is generated in-vivo through the diffusion-controlled reaction between nitric oxide and superoxide. Peroxynitrite has been linked to nitration of protein and DNA as well as to DNA strand breaks. Accumulation of mutations and/or other kinds of DNA damage represent a carcinogenic risk. The accurate measurement of peroxynitrite concentration has been a challenge since this analyte is very unstable and reacts with many cellular targets. Development of analytical techniques capable of rapid and sensitive detection of this fast-reacting and damaging agent …

Profiling Cell Surface Sialylation And Desialylation Dynamics Of Immune Cells, Dan Wang

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Sialic acids (SAs) are a diverse family of naturally occurring 2-keto-3-deoxy-nononic acids that are involved in a wide range of biological processes, including early fetal development, cellular recognition, and utilization by microbes. While it is clear that cell surface SAs are highly involved in the immune system, the sialylation status of individual immune cells and functions are still unknown. In this study, I combined the newly developed LC-MS/MS methods with flow cytometry and confocal microscopy to systematically study the sialylation and desialylation dynamics of macrophages at different conditions.

First, I developed an accurate LC-MS/MS method to quantify free SA in …

Development And Validation Of Uplc/Ms/Ms Methods For Quantification Of Gangliosides In The Clinical Study Of Ganglioside Gm3 Synthase Deficiency, Qianyang Huang

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Gangliosides are a large subclass of glycosphingolipids that structurally characterized by the addition of mono- or poly-sialyated carbohydrate moieties onto the ceramide scaffold. They are presented at significant abundance in the central nervous system of vertebrates. Their interactions with trans-membrane receptors, especially receptor tyrosine kinase, are believed to play critical roles in modulating signal transduction events during cell proliferation, differentiation, migration, and adhesion. Furthermore, the disruption and dysfunction on their metabolism have been found to be associated with various neurodegenerative disorders, such as Alzheimer disease, Parkinson’s disease, and GM3 synthase deficiency. GM3 synthase deficiency (GSD) is a newly identified neurological …

Study Of Electron Transfer Through The Reductase Domain Of Neuronal Nitric Oxide Synthase And Development Of Bacterial Nitric Oxide Synthase Inhibitors, Yue Dai

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Crystal structure of neuronal Nitric Oxide Synthase reductase (nNOSr) implies that large-scale domain motion is essential for electron transfer. However, the details are not well understood. To address this, we generated a functioning “Cys-lite” version of nNOSr and then replaced the nNOSr Glu816 and Arg1229 residues with Cys in the FMN and FAD domains (CL5SS) in order to allow cross-domain disulfide bond formation under pH 9 or to cross-linking using bis-maleimides. Cross-linked CL5SS exhibited a =95% decrease in cytochrome c reductase activity and reduction of the disulfide bond restored the activities. The results demonstrate that a conformational equilibrium involving FMN …

The Role Of Rnase L In Type I Diabetes And Development Of Quantitative Lc-Ms/Ms Methods For The Pharmacological Studies Of Anti-Cancer Drug Candidates, Chun Zeng

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PROJECT I: The cause of type I diabetes continues to be a focus of investigation. In this study, we found that 2-5A dependent RNase L (RNase L), an IFN-a-inducible enzyme that functions in IFN action against viruses and cell proliferation, played an important role in dsRNA-induced onset of type I diabetes. By using RNase L deficient RIP-B7.1 mice which are more vulnerable to environmental harmful factors such as viral infection, we demonstrated that deficiency of RNase L in mice resulted in a significant delay of diabetes onset induced by polyinosinic:polycytidylic acid (poly I:C), a type of dsRNA, and streptozotocin (STZ). …

Application Of Advanced Analytical Technologies To Drug Development Studies And Cancer Detection, Ramakrishna Reddy Voggu

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In the past decade, bioanalytical method development has become an integral part of the clinical diagnosis, biomarker discovery, and drug discovery and development. The new and emerged bioanalytical techniques allow the quantitative and qualitative analysis of bio molecules with remarkably high sensitivity and specificity. Specifically, these bioanalytical methods based on LC-MS and methylation-specific PCR are well suited for detecting low-abundance metabolites in various biological fluids and DNA in plasma and tissues for biomarker investigation. They offer great clinical promise for early disease diagnosis and therapeutic intervention. This dissertation broadly organized into two parts, part one talks about the application of …

Characterization Of Cold Drawn Poly L Lactic Acid By Raman Spectroscopy And Raman Hyperspectral Imaging, Ramya Sudha Muttavarapu

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Artificial modulation of bone growth and repair processes is an important area of current research. Studies have established that the micro mechanical and bulk physical properties of a polymer implant influence its performance behavior and degradation patterns in the body. One important parameter for the class of polymers known as polylactides is the degree of crystalline character. Hence, studying the bulk and micro crystalline composition of an important polymer implant, Poly-L-Lactic acid is the objective of this study. Poly-L-Lactic acid has elastic properties that can be tailored to resemble natural bone and its reaction to mechanical stress. Varying degrees of …

The Role Of Platelets In Hyaluronan Degradation, Sami Albeiroti

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Following tissue injury, platelets rapidly interact with the exposed extracellular matrix (ECM) of blood vessel wall and the surrounding tissues. Hyaluronan (HA) is a major polysaccharide component of the ECM and plays a significant role in regulating inflammation. Human platelets can degrade HA from the surfaces of activated endothelial cells into fragments capable of inducing immune responses by monocytes. In addition, platelets contain the enzyme hyaluronidse-2 (HYAL2), one of two major hyaluronidases that digest HA in somatic tissues. The deposition of HA increases in the inflamed tissues in several inflammatory diseases, including Inflammatory Bowel Disease (IBD). This study provides evidence …