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A Molecular Simulation Study Of Antibody-Antigen Interactions On Surfaces For The Rational Design Of Next-Generation Antibody Microarrays, Derek B. Bush

Theses and Dissertations

Antibody microarrays constitute a next-generation sensing platform that has the potential to revolutionize the way that molecular detection is conducted in many scientific fields. Unfortunately, current technologies have not found mainstream use because of reliability problems that undermine trust in their results. Although several factors are involved, it is believed that undesirable protein interactions with the array surface are a fundamental source of problems where little detail about the molecular-level biophysics are known. A better understanding of antibody stability and antibody-antigen binding on the array surface is needed to improve microarray technology. Despite the availability of many laboratory methods for …

System Genetic Analysis Of Mechanisms Underlying Excessive Alcohol Consumption, Maren L. Smith

Theses and Dissertations

Increased alcohol consumption over time is one of the characteristic symptoms of Alcohol Use Disorder (AUD). The molecular mechanisms underlying this escalation in intake is still the subject of study. However, the mesocortical and mesolimbic dopamine pathways, and the extended amygdala, because of their involvement in reward and reinforcement are believed to play key roles in these behavioral changes. Multiple gene expression studies have shown that alcohol affects the expression of thousands of genes in the brain. The studies discussed in this document use the systems biology technique of co-expression network analysis to attempt to find

patterns within genome-wide expression …

A Weighted Gene Co-Expression Network Analysis For Streptococcus Sanguinis Microarray Experiments, Erik C. Dvergsten

Theses and Dissertations

Streptococcus sanguinis is a gram-positive, non-motile bacterium native to human mouths. It is the primary cause of endocarditis and is also responsible for tooth decay. Two-component systems (TCSs) are commonly found in bacteria. In response to environmental signals, TCSs may regulate the expression of virulence factor genes.

Gene co-expression networks are exploratory tools used to analyze system-level gene functionality. A gene co-expression network consists of gene expression profiles represented as nodes and gene connections, which occur if two genes are significantly co-expressed. An adjacency function transforms the similarity matrix containing co-expression similarities into the adjacency matrix containing connection strengths. Gene …

Meta-Analysis Of Microarray Data To Assess Gender Biased Differential Gene Expression In Hepatic Tissue, Amira Salah Eldin Mahmoud Elbakry

Theses and Dissertations

Hepatocellular carcinoma (HCC) is the second deadliest cancer globally, and with an estimated 782,000 new cases in 2012, it is the fifth most common cancer in men and ninth in women. HCC is of particular concern in Egypt because of the high prevalence of Hepatitis C Virus (HCV). Due to its poor prognosis, HCC is the leading cause of cancer-related deaths in Egypt. A gender disparity is observed in liver cancer cases, with higher prevalence in men by three to five fold. This sex bias is even more pronounced in mouse models of HCC, which was found to be sex …

Investigating The Role Of The Synaptic Transcriptome In Ethanol-Responsive Behaviors, Megan A. O'Brien

Theses and Dissertations

Alcoholism is a complex neurological disorder characterized by loss of control in limiting intake, compulsion to seek and imbibe ethanol, and chronic craving and relapse. It is suggested that the characteristic behaviors associated with the escalation of drug use are caused by long-term molecular adaptations precipitated by the drug’s continual administration. These lasting activity-dependent changes that underlie addiction-associated behavior are thought, in part, to depend on new protein synthesis and remodeling at the synapses. It is well established that mRNA can be transported to neuronal distal processes, where it can undergo localized translation that is regulated in a spatially restricted …

Characterization Of A Novel Ring-Type Ubiquitin E3 Ligase Ghring2 Involved In Cotton Fiber Development, Shiva Theja Reddy Soma

Theses and Dissertations

The ubiquitin-proteasome proteolysis pathway is responsible for the degradation of abnormal and short-lived proteins to regulate many important biochemical activities in eukaryotes. By employing affymetrix microarray analysis, we have identified a novel ubiquitin ligase E3 gene GhRING2 that is expressed differentially between two G. hirsutum lines - Texas Marker-1 and Chromosome Substitution Line CS-B25. The complete GhRING2 gene sequence was obtained by genomic and cDNA walking. The expression of GhRING2 in cotton fiber is developmentally regulated, suggesting that the ubiquitin-proteasome pathway may regulate cotton fiber growth and development. Using a yeast two-hybrid assay GhRING2 was found to interact with a …

Molecular Modeling Of Dna For A Mechanistic Understanding Of Hybridization, Terry Jacob Schmitt

Theses and Dissertations

DNA microarrays are a potentially disruptive technology in the medical field, but their use in such settings is limited by poor reliability. Microarrays work on the principle of hybridization and can only be as reliable as this process is robust, yet little is known at the molecular level about how the surface affects the hybridization process. This work uses advanced molecular simulation techniques and an experimentally-parameterized coarse-grain model to determine the mechanism by which hybridization occurs on surfaces and to identify key factors that influence the accuracy of DNA microarrays. Comparing behavior in the bulk and on the surface showed, …

Analyzing The Functions Of Human Polynucleotide Phosphorylase (Hpnpaseold-35), Upneet K. Sokhi

Theses and Dissertations

RNA degradation plays a fundamental role in maintaining cellular homeostasis, along with being a part of normal regulatory mechanisms, whether it occurs as a surveillance mechanism eliminating aberrant mRNAs or during RNA processing to generate mature transcripts. 3’-5’ exoribonucleases are essential mediators of RNA decay pathways, and one such evolutionarily conserved enzyme is polynucleotide phosphorylase (PNPase). The human homologue of this fascinating enzymatic protein (hPNPaseold-35) was cloned a decade ago in the context of terminal differentiation and senescence through a novel ‘overlapping pathway screening’ approach. Since then, significant insights have been garnered about this exoribonuclease and its repertoire of expanding …

Identification And Characterization Of Ethanol Responsive Genes In Acute Ethanol Behaviors In Caenorhabditis Elegans, Joseph Alaimo

Theses and Dissertations

Alcohol abuse and dependence are complex disorders that are influenced by many genetic and environmental factors. Acute behavioral responses to ethanol have predictive value for determining an individual’s long-term susceptibility to alcohol abuse and dependence. These behavioral responses are strongly influenced by genetics. Here, we have explored the role of genetic influences on acute behavioral responses to ethanol using the nematode worm, Caenorhabditis elegans. First, we explored the role of ethanol metabolism in acute behavior responses to ethanol. Natural variation in human ethanol metabolism machinery is one of the most reported and reproducible associations found to alter drinking behavior. Ethanol …

The Effects Of Mismatches And Probe Tethering Configurations On The Stability Of Dna Duplexes On Surfaces, Kyle Evan Pratt

Theses and Dissertations

DNA microarrays are chip-based, analysis tools which can perform hundreds of thousands of parallel assays to determine the identity of genes or gene expression levels present in a sample. They have been identified as a key technology in genomic sciences and emergent medical techniques; however, despite their abundant use in research laboratories, microarrays have not been used in the clinical setting to the fullest potential due to the difficulty of obtaining reproducible results. Microarrays work on the principle of DNA hybridization, and can only be as accurate as this process is robust. Fundamental, molecular-level understanding of hybridization on surfaces is …

Identification Of Virulence Factors In Edwardsiella Ictaluri, Jingjun Lu

Theses and Dissertations

Edwardsiella ictaluri is the causative agent of enteric septicemia of catfish (ESC), which is one of the most important diseases impacting the US catfish industry. Though this disease has been very common, progress has been slow to find an economical and practical treatment method. Our long-term goal is to determine the mechanisms of E. ictaluri virulence in ESC. The overall objective of this study was to identify E. ictaluri genes required for host encounter and serum resistance and to determine their roles in pathogenesis. The central hypothesis is that E. ictaluri must differentially regulate its genes to invade fish and …

The Role Of Host-Tumor Interactions In Liver Metastasis Of Colorectal Cancer, Yu Zhang

Theses and Dissertations

Colon cancer is the third most frequent cancer and the second leading cause of cancer deaths in the United States. Liver metastasis is the major cause of death in colon cancer. Successful metastases depend on productive collaborations between tumor cells and host-derived cells in the tumor microenvironment, target organ environments, and cells in the hematopoietic compartment.

To identify the host-tumor interactions promoting liver metastasis and their molecular and cellular mediators, an orthotopic mouse model of liver metastasis of colon cancer was established that recapitulates all stages of tumor growth and metastasis. A highly metastatic mouse carcinoma cell line CT26-FL3 was …

The Anti-Angiogenic Effects Of Sparstolonin B, Henry Rhodes Bateman

Theses and Dissertations

Sparstolonin B (SsnB) is a novel bioreactive compound isolated from Sparganium stoloniferum, an herb historically used in Traditional Chinese Medicine as an anti-tumor agent. SsnB has demonstrated anti-inflammatory properties, inhibiting Toll-like receptor mediated inflammation in isolated macrophages and in mice. Angiogenesis, the process of new capillary formation from existing blood vessels, is dysregulated in many pathological disorders, including atherosclerosis, diabetic retinopathy, and tumor growth. The goal of the project was to investigate the anti-angiogenic effects of SsnB.

The first part of the project utilized in vitro functional assays to study how SsnB affected endothelial cells. SsnB inhibited endothelial cell tube …

The Role Of P62 In Osteoclastogenesis And Paget’S Disease Of Bone, Tamer Hadi

Theses and Dissertations

Paget’s disease (PDB) is the second most common metabolic bone disease after osteoporosis, affecting up to 3% of adults over age 55. It is characterized by focal lesions of bone resorbed by hyperactive osteoclasts coupled with rapid formation of highly disorganized, low quality bone formed by osteoblasts. Such lesions cause skeletal deformity, fractures, and other symptoms that significantly decrease quality of life. In 2001, mutations in the SQSTM1/p62 gene were found in a subset of Paget’s patients. The work summarized in this dissertation sought to answer two broad questions: what is the function of p62 in normal bone homeostasis and …

Molecular Pathways Involved In Calcineurin Inhibitor Nephrotoxicity In Kidney Allograft Transplants, Huong Nguyen

Theses and Dissertations

ABSTRACT MOLECULAR MECHANISMS AND GENE SIGNATURES INVOVLED IN CALCINEURIN INHIBITOR NEPHROTOXICITY IN KIDNEY ALLOGRAFT By Huong Le Diem Nguyen, M.S. A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Physiology at Virginia Commonwealth University. Virginia Commonwealth University, 2011. Major Director: Valeria Mas, Ph.D. Associate Professor, Department of Surgery and Pathology Director of Molecular Transplant Research Laboratory, Division of Transplant Calcineurin inhibitors (CNI), cyclosporin A and tacrolimus, are potent immunosuppressive agents but induce toxicities causing damages and graft dysfunction, and have been suggested to contribute to late-term loss of graft in kidney transplant …

Identification Of Genes Differentially Expressed In Elongating Fiber In A Cotton Chromosome Substitution Line Cs-B25, Samuel Sunil Kumar Bandi

Theses and Dissertations

Identification of genes specific to fiber development would improve the efforts in developing cotton plants with superior fiber quality. Through genetic introgression, 17 interspecific chromosome substitution lines (CS-B lines) of upland cotton in G. hirsutum (TM-1) have been developed and released recently. These substitution lines have TM1 as background and contain either whole chromosome or chromosome arms of G. barbadense (line 3-79) chromosomes. CS-B25 has chromosome 25 from G. barbadense substituted into TM-1 G. hirsutum was reported to show superior fiber properties. In this study, suppression subtraction hybridization (SSH) combined with Affymetrix cotton genome microarray arrays were used to identify …

Mosaicism For Trisomy21: Utility Of Array-Based Technology For Its Detection And Its Influence On Telomere Length And The Frequency Of Acquired Chromosome Abnormalities, Chariyawan Charalsawadi

Theses and Dissertations

The primary aim of this study was to determine the effectiveness of array-based technology for detecting and quantifying the presence of mosaicism. This aim was achieved by studying individuals having mosaicism for Down syndrome. SNP arrays were performed on 13 samples from individuals with mosaicism for trisomy 21, 13 samples from individuals with normal chromosome 21complements (negative controls) and 5 samples from individuals with full or partial trisomy 21 (positive controls). In addition, BAC arrays were processed on 6 samples from individuals with mosaicism for trisomy 21, 3 negative controls and 1 positive control. These studies have shown that array-based …

Identification Of Micrornas In Bovine Spermatozoa With Implications Of Fertility, Lashonda S (Lashonda Shakita) Robertson

Theses and Dissertations

MicroRNAs are small RNA molecules that could possibly play a major role in fertility. In the experiment, spermatozoa were extracted from bovine followed by an extraction of total RNA. Bovine spermatozoa were extracted from two bulls of different fertility, high and low fertility. An expression array was done to compare the expression levels of the microRNAs. It was shown that thousands of microRNAs are present in bovine spermatozoa but only a small amount was significantly expressed. The microRNAs from low fertility bulls were more highly expressed than those in high fertility bulls. A Bioanalyzer gel was used to confirm the …

Improving Structural And Functional Annotation Of The Chicken Genome, Teresia Buza

Theses and Dissertations

Chicken is an important non-mammalian vertebrate model organism for biomedical research, especially for vaccine production and the study of embryology and development. Chicken is also an important agricultural species and major food source for high-quality protein worldwide. In addition, chicken is an important model organism for comparative and evolution genomics. Exploitation of this genome as a biomedical model is hindered by its incomplete structural and functional annotation. This incomplete annotation makes it difficult for researchers to model their functional genomics datasets. Improving structural and functional annotation of the chicken genome will allow researchers to derive biological meaning from their functional …

Genetic Dissection Of Triterpenoid Saponin Production In Chenopodium Quinoa Using Microarray Analysis, Derrick James Reynolds

Theses and Dissertations

Quinoa (Chenopodium quinoa Willd.) is an important food crop for subsistence farmers in the Altiplano (high plains) of Peru, Bolivia, and Argentina. Saponins are part of a diverse family of secondary metabolites that are found in high concentrations in the pericarp of many varieties of quinoa. Due to their bitter taste and anti-nutritive properties, saponins must be removed before the quinoa grain is consumed. There are ‘sweet’ varieties of quinoa that have significantly reduced levels of saponin. Previous research suggests saponin production is controlled by a single locus. The major objective of this research was to elucidate the genetic …

Probe Level Analysis Of Affymetrix Microarray Data, Richard Ellis Kennedy

Theses and Dissertations

The analysis of Affymetrix GeneChip® data is a complex, multistep process. Most often, methodscondense the multiple probe level intensities into single probeset level measures (such as RobustMulti-chip Average (RMA), dChip and Microarray Suite version 5.0 (MAS5)), which are thenfollowed by application of statistical tests to determine which genes are differentially expressed. An alternative approach is a probe-level analysis, which tests for differential expression directly using the probe-level data. Probe-level models offer the potential advantage of more accurately capturing sources of variation in microarray experiments. However, this has not been thoroughly investigated, since current research efforts have largely focused on the …

Maternal High-Salt Diet During Pregnancy In Sprague Dawley Rats Programs Exaggerated Stress-Induced Blood Pressure And Heart Rate Responses In Adult Female Offspring, Summer Hayes King

Theses and Dissertations

The prenatal environment has been shown to have lasting effects on cardiovascular health. In the present study, pregnant rats were fed a 0.7% NaCl normal salt (NS) diet or an 8% NaCl high salt (HS) diet throughout pregnancy. Adult offspring were fitted with radiotelemetry probes to continuously measure blood pressure and heart rate. Rats were placed in restraining cages to test for a programmed acute stress hyperresponsiveness. Offspring were challenged with HS diet for one week to determine if blood pressure salt sensitivity had been programmed by the prenatal HS diet. Animals were killed following resting and acute stress conditions, …

Development Of Informative Priors In Microarray Studies, Kassandra M. Fronczyk

Theses and Dissertations

Microarrays measure the abundance of DNA transcripts for thousands of gene sequences, simultaneously facilitating genomic comparisons across tissue types or disease status. These experiments are used to understand fundamental aspects of growth and development and to explore the underlying genetic causes of many diseases. The data from most microarray studies are found in open-access online databases. Bayesian models are ideal for the analysis of microarray data because of their ability to integrate prior information; however, most current Bayesian analyses use empirical or flat priors. We present a Perl script to build an informative prior by mining online databases for similar …

Molecular-Genetic And Structural Analyses Of The Nifhdkx Proteins Of The Nitrogenase System, Surobhi Lahiri

Theses and Dissertations

The nitrogenase enzyme is the biochemical machiner responsible for the conversion of the largely unavailable nitrogen to the easily assimilable ammonia for living organisms by the process termed as biological nitrogen fixation (BNF). This study was focused on understanding the various structural and functional aspects of the nitrogenase enzyme related to maturation and assembly of the FeMo-cofactor (FeMoco) metallocluster of the MoFe protein (the site for final substrate reduction), development of a dimeric MoFe protein and the structural homology of nitrogenase with other metalloenzymes. This research was specifically directed towards the NifHDKX proteins in which the nifHDK genes are the …

A Comparison Of Microarray Analyses: A Mixed Models Approach Versus The Significance Analysis Of Microarrays, Nathan Wallace Stephens

Theses and Dissertations

DNA microarrays are a relatively new technology for assessing the expression levels of thousands of genes simultaneously. Researchers hope to find genes that are differentially expressed by hybridizing cDNA from known treatment sources with various genes spotted on the microarrays. The large number of tests involved in analyzing microarrays has raised new questions in multiple testing. Several approaches for identifying differentially expressed genes have been proposed. This paper considers two: (1) a mixed models approach, and (2) the Signiffcance Analysis of Microarrays.

Design And Development Of Oligonucleotide Microarrays And Their Application In Diagnostic And Prognostic Estimation Of Human Gliomas, G. Scott Taylor

Theses and Dissertations

DNA microarrays represent an ultra-high throughput gene expression assay employed to study the transcriptomic profiles of biological tissues. These devices are increasingly being used to study many aspects of gene regulation, and there is growing interest in the biotechnology and pharmaceutical industries for developing such devices in efforts toward rational product/drug design. The DNA microarray also provides a unique and objective means for diagnosis and prognosis of human diseases based on patterns of gene expression. This is especially important in cancer research and the thrust toward personalized medicine. This dissertation details the design and development of oligonucleotide microarrays and the …