Molecular Biology Commons^™

Transcriptomics Of Chicken Primordial Germ Cells, Nhung Thi Nguyen

Graduate Theses and Dissertations

Chicken primordial germ cells (PGCs) are derived from extraembryonic tissue of the embryo and first appear at stage X of development. They enter the bloodstream and migrate to the genital ridge, unite with somatic tissue to form a developing gonad, and then differentiate to sperm or ova (Fujimoto et al., 1976). Understanding molecular features of both male and female PGCs not only clarify the differentiation mechanism of such cells toward different germ lines, but will also help in selecting for highly productive types of commercial chicken. Most previous studies focused on the location of PGCs (Eyal-Giladi et al., 1981; Swift …

Autophagy And Its Potential Role In Stress And Feed Efficiency Using Avian Lines, Alissa Laura Piekarski

Graduate Theses and Dissertations

Autophagy is a highly conserved cellular mechanism that is responsible for the degradation and recycling of damaged organelles. Recently, autophagy has been involved in critical roles during overall development of the organism and degradation of damaged cellular components. This pathway has witnessed dramatic growth in the last few years and has been extensively studied in yeast and mammals, however, there is a paucity of information in avian (non-mammalian) species. First, we characterized genes involved in the autophagy pathway in male and female Jungle Fowl to determine gender and tissue specific differences. Secondly, tissue and genotype differences in Japanese quail selected …

In Vivo Periodical Monitoring Of Immune Cell Infiltration In Response To Feathers And Intramuscular Injection Of Ionps Using The Pulp (Dermis) Of Growing Feathers As Test Site Tissue In Chickens, Olfat Taleb Alaamri

Graduate Theses and Dissertations

The applications of nanoparticles are growing, but little is known about their interactions with the immune system as most studies did not use suitable in vivo test systems. The purpose of these studies was to investigate the utility of chicken growing feathers as an in vivo test site for iron oxide nanoparticle (IONP) interactions with the immune system. The first objective of this study was to monitor leukocyte infiltration into the growing feather pulp upon the administration of IONP and IO-mIgG preparations. The second objective was to test the utility of IONPs as vaccine adjuvants by monitoring primary and memory …

Understanding The Causal Agent Of Rose Rosette Disease, Patrick Louis Di Bello

Graduate Theses and Dissertations

A number viruses are known to infect roses, ranging from those in the genera Nepovirus, and Ilarvirus, which have been reported since the inception of rose virology, to recently discovered viruses in the genera Carmovirus, Closterovirus, Emaravirus, Luteovirus, Rosadnavirus, and Potyvirus. Of the viral diseases in rose, arguably the most damaging is Rose rosette (RRD), which is associated with the Emaravirus, Rose rosette virus (RRV). The objective of this thesis is to fill in the gaps in knowledge on the epidemiological aspects of RRD and RRV. There has been significant progress in the epidemiology of the RRD agent prior to …

Susceptibility Of Parkinson’S Disease Following Mild Blast Traumatic Brain Injury, Glen Howel Galicia Acosta

Open Access Theses

Blast injury-induced neurotrauma (BINT) is steadily increasing in prevalence due to escalated terror activity and constitutes the signature injury associated with current military conflicts. BINT produces significant neurological deficiencies and there is a growing concern that the injury may produce long-term consequences that affect the resilience and the performance of soldiers. One of the potential consequences is an increased susceptibility to Parkinson's disease (PD). A vital goal aimed at curtailing the post-deployment long-term consequences of blast injury-induced neurotrauma is to further our knowledge of pathogenic mechanisms responsible for the escalation of post injury diseases. The purpose of this project is …

Characterization Of The Effects Of Bfgf And Gsk3Β Inhibitors On Embryoid Bodies Derived From Human Pluripotent Stem Cells, Jonathan Earls

Graduate Theses and Dissertations

Embryoid body (EB) formation is a common first step in many human pluripotent stem cell (hPSC) differentiation protocols. Previous work suggests that EBs are sensitive to growth factor withdrawal if they are derived from hPSCs maintained in feeder independent media such as mTeSR1. To promote cell survival, EBs generated from mTeSR1-adapted hPSCs are sometimes cultured in a medium that contains basic fibroblast growth factor (bFGF), a trophic factor often used in hPSC cultures to maintain self-renewal. This distinguishes feeder independent hPSCs from feeder dependent hPSCs. The purpose of this study was to characterize the effects of bFGF as well as …

Comparison Of Different Modulators That Affect Macrophage Activation In Vitro, Alda Alexa Díaz Pérez

Graduate Theses and Dissertations

Inflammation is known as a mechanism to regulate and control infections as well as promote tissue repair. Macrophages (Mф) are known to be a major cell type in the initiation, sustainability and resolution of inflammation. Moreover, Mф are essential for the remodeling process that is also known as the wound healing response. The objective of this research was to compare five modulators (acetylsalicylic acid (ASA), dexamethasone (DEX), prostaglandin E2 (PGE2), iloprost, and resolvin D1 (RvD1) for their anti-inflammatory effects on macrophages in vitro. Then, Mф phenotype in terms of gene expression and secreted cytokine response was determined. Our study compared …

A Morpho-Physiological Analysis Of Diverse Drought Resistant And Sensitive Rice Genotypes To Identify Distinguishing Anatomical Root Phenes, Clinton Philip Greub

Graduate Theses and Dissertations

A set of 37 genotypes selected from a survey of over 200 diverse germplasms, for their diverse drought tolerance mechanisms, were analyzed in this study for root physiological traits that are indicative of different drought resistance mechanisms. The genotypes include controls such as the drought sensitive IR64 and Nipponbare, along with the drought resistant Nagina 22 (N22) and Bengal landraces to identify distinguishing phenotypic features of resistant accessions. Anatomical traits studied include total areas of the cross-sections of the stele, cortex, aerenchyma, cortical-cell, and xylem tissue; the number of aerenchyma, xylem vessels, cortical cell file number and size that together …

Production And Characterization Of Islet Amyloid Polypeptide Using Recombinant Technology, Emilio Duverna

Graduate Theses and Dissertations

Islet amyloid polypeptide (IAPP) also called amylin is an amyloid-forming protein; IAPP is a proteinaceous hormone that comprises 37 amino acid residues. It is secreted along with insulin from the pancreatic β-cells to help it regulate the uptake and removal of glucose in the bloodstream. IAPP has been observed in the amyloid deposits found in pancreatic β-cells of most patients suffering from type II diabetes mellitus. This research project aims at producing recombinant amylin peptide. To achieve this goal, we used the pBAD plasmid vector which we introduced into Escherichia coli to express the peptide. Although the vector was successfully …

Functional Significance Of Gill Claudin Proteins In Rainbow Trout (Oncorhynchus Mykiss) Osmoregulation, Joanna Katarzyna Bujak

Graduate Theses and Dissertations

Claudin proteins, a key element of tight junction complexes, are known to control paracellular permeability. In euryhaline fish, changes in claudin abundance and localization are critical during salinity acclimation. In seawater, a leaky paracellular pathway that facilitates sodium extrusion is hypothesized to be controlled by claudin proteins. The aim of this study was to evaluate the role of Claudin-10c, -10d -10e and Claudin-30 in gill function in freshwater (FW) and seawater (SW) rainbow trout (Oncorhynchus mykiss). I examined mRNA and protein abundance along with cellular localization. A tissue distribution survey showed that all of the claudins studied were predominantly expressed …

Impacts Of Micrornas On Skeletal Muscle Protein Synthesis And Mitochondrial Quality, David Lee

Graduate Theses and Dissertations

microRNA (miRNA) post-transcriptional modification is becoming a well-established mechanism for controlling mRNA translation. microRNAs -1, -133, and -206 are under the control of skeletal muscle promoters and affect muscle plasticity and metabolic health. A detailed review on the generation and processing of miRNAs with a view to skeletal muscle brings up intriguing connections in the transcriptional connections between multiple miRNAs. Additionally, exciting new research has defined a role of miRNAs in skeletal muscle mitochondria showing an additional, direct link to metabolic function. Multiple investigations in models of exercise, aging, hypertrophy, and injury have shown how these interventions can affect miRNA …

Local Modulation And Measurement Of Macrophage-Derived Bioactive Proteins From Implanted Biomaterials In Rat, Geetika Bajpai

Graduate Theses and Dissertations

Fibrosis around the implanted medical devices is a severe problem that can plague long-term device reliability. Activation of macrophage phenotype (macrophage polarization) has emerged as a new and possible means for reducing fibrosis in the fields of biomaterials and regenerative medicine. Macrophages are phagocytic cells that respond to microenvironmental cues that direct their phenotype. Macrophage activation has been widely studied in mouse and human in the context of tumor biology, yet little information is available regarding how macrophage activation could be used in a biomaterials context. Further, rats rather than mice are the common subjects in biomaterials experiments. A significant …

New Strategies To Reveal Protein Candidates In Protein-Protein Interactome Study, Meng-Chieh Chen

Open Access Theses

Comprehensive protein-protein interaction network analysis can help reveal protein functions in a system-wide manner. A reliable knowledgebase of interaction networks is not only important for selecting the candidates for drug therapies, but also for evaluating the disease risk. In current interaction databases, 322579 interactions comprised of 56460 proteins have been reported (statistical analysis from APID: Agile Protein Interaction DataAnalyzer; http://bioinfow.dep.usal.es/apid/index.htm). The huge datasets are contributed mainly by yeast -two -hybrid (Y2H) screening and affinity-purification followed by mass spectrometry (AP-MS). High false positive rates and failing to cover certain interaction categories are the limitations of these two methods. Here, we developed …

1,25-Dihydroxyvitamin D Alters Lipid Metabolism And Epithelialto- Mesenchymal Transition In Metastatic Epithelial Breast Cancer Cells, Alle Nicole Barnard

Open Access Theses

Evidence suggests that high vitamin D status (marked by serum 25-hydroxyvitamin D, 25(OH)2 D) is associated with a decreased risk of breast cancer. It has been established that 1,25-dihydroxyvitamin D (1,25(OH) 2 D) can alter glycolysis and the Krebs cycle of breast cancer cells (Jiang et al., 2010; Zheng et al., 2013) but little information is available on 1,25(OH) 2 D's alterations of lipid metabolism in breast cancer cells. Thus, the current research investigates if there was an effect of 1,25(OH) 2 D on proteins that regulate lipid metabolism in MCF10A, MCF10A-ras, MCF10CA1h, and MCF10CA1a epithelial breast cancer cells. While …

Neurotrophins And Their Effects On Breast Cancer Cell Proliferation And Migration, Kayla Elise Minser

Open Access Theses

Cancer is a large health issue in all parts of the world. In the United States alone, approximately 1 in 4 deaths are cancer related. Breast cancer is a particularly prevalent form, accounting for a little over 14 percent of all cancer incidence. The largest obstacle to overcome for breast cancer morbidity is metastasis. Over 90 percent of all breast cancer related deaths are due to metastasis. Because metastasis is a complex, multi-step process, it is difficult to treat. A recent observation in the Kirshner lab has revealed a type of phenotypic plasticity, where migratory cancer cells have a neuronal-like …

Pharmaceuticals And Personal Care Products: Emerging Contaminants In Aquatic Ecosystems, Jenny E. Zenobio

Open Access Theses

In recent years, the presence of pharmaceuticals and personal care products (PPCPs) in aquatic systems has led to research on their fate and effects. PPCPs have been found in mixture in wastewater effluents, surface, ground, and drinking water at low concentrations from areas of intense urbanization. Although adverse effects to human health from the current environmental concentrations are unlikely, the impacts to ecological receptors are not clear. We performed field and laboratory studies to quantify and evaluate effects of PPCPs on fish. First, a field study was conducted at the Baca National Wildlife Refuge, Colorado (2010-2012) because a portion of …

Development Of Novel Subunit Vaccine Against H5n1 Influenza, Lu Zhang

Graduate Theses and Dissertations

Influenza is a common infectious disease resulting from a frequently mutated RNA virus. Vaccination is currently the most effective method to prevent people from seasonal or pandemic influenza. The production of traditional egg-based influenza vaccine is time-consuming and provides limited effect against new strains. Therefore, it is necessary to develop a rapid method to produce influenza vaccines. We proposed a novel influenza vaccine based on the E.coli expression system. Hemagglutinin (HA) is the major target surface protein of influenza virus for vaccine development. In this study, we sub-cloned the HAs encoding gene into an E. coli expression vector; the signal …

The Development Of An In Vivo Microdialysis Collection Method Of Cytokines From Brain Tissue, Anthony W. Herbaugh

Graduate Theses and Dissertations

In this thesis, different methods to improve the microdialysis collection procedure for cytokines from brain tissue are presented. The first method was based on stopped flow and results indicating that no significant difference in relative recovery between stopped flow and continuous flow are shown. The second method is an antibody bead-based enhancement method. With the antibody bead-based method, a 3.5 fold increase in the collected concentrations of Chemokine (C-C motif) Ligand 2 (CCL2) were observed. However, there was no significant increase in the in vivo collection efficiency of Interleukin-6 (IL-6) using the antibody enhancement. Finally the development of an in-house …

The Role Of Histone H3 And H4 In Centromere Function And Genome Integrity, Payel Chaudhuri

Graduate Theses and Dissertations

Histone H2A plays an important role in chromosomal segregation among parent and daughter cells during mitosis. While it is established that this histone is important in maintaining chromosome number in cell, further work is carried out to explore the role of other histones like H3 and H4 for similar effects. A systematic study is initiated by screening a library based on mutation of different amino acid residues in these histones. This detailed screening identified specific regions within H3 and H4, which are critically important for centromeric function. These histones residing near the DNA entry/exit region of nucleosome effects the functionality …

Molecular And Biochemical Mechanisms Of Pathogenesis In The Maize Foliar Pathogen Cercospora Zeae-Maydis, Winfred-Peck Dorleku

Graduate Theses and Dissertations

GLS is a serious foliar disease of maize, a major staple crop grown commercially in the USA for both human and animal feed production, and increasingly, for ethanol production. The disease is caused by two Cercospora species, C. zeae-maydis and C. zeina, both of which infect maize in the USA and in other parts of the world, with yield losses potentially greater than 50%, depending on local conditions. In culture, C. zeae-maydis produces a phytotoxic, host non-specific perylenequinone, cercosporin, and abscisic acid (ABA), for which there is no known pathological or physiological function in the fungus. Experimental evidence indicates …

Characterization Of Caxck31, A Bacterial Calcium/Proton Antiporter, Marc Robert Ridilla

Open Access Dissertations

To better understand a class of transporters known as Calcium/Cation Antiporters (CaCAs), the bacterial calcium/proton antiporter CAXCK31 was purified and characterized. New methods were developed for its heterologous overexpression and purification. These methods help to define stress responses to toxic membrane overproduction in E. coli and may be broadly applicable to studies of membrane proteins. The results from a variety of biochemical and biophysical experiments demonstrated that CAXCK31 exists as a dimer in the membrane and can be purified in the dimeric state. The methods used include chemical cross-linking, FRET, and SEC-MALS. In addition, various transport properties of CAXCK31, including …

Structural Studies On The Rubella Virus Capsid Protein And Its Organization In The Virion, Vidya Mangala Prasad

Open Access Dissertations

Rubella virus is a leading cause of birth defects due to infectious agents. When contracted during pregnancy, rubella infection leads to severe damage in fetuses. Despite its medical importance, very little is known about the structure of the pleomorphic rubella virus as compared to its alphavirus relatives. The rubella capsid protein is a critical structural component of virions as well as a key factor in virus-host interactions. Three crystal structures of the structural domain of the rubella capsid protein have been described here. The polypeptide fold of the capsid protomer has not been observed previously. The capsid protein structure, along …

The Termite Digestome: Understanding The Digestive Physiology Involved In Lignocellulosic Biomass Degradation, Zachary John Karl

Open Access Dissertations

The purpose of this research was to advance the understanding of lower termite digestive physiology and discover potential biocatalysts that can aid in the degradation of lignocellulosic biomass. Various protein characterization and gene expression methods were used throughout this research in order to accomplish these objectives. The results of this dissertation indicate that: 1) termites and their symbionts act in a synergistic manner to degrade biomass in vitro, 2) the host fraction of the gut (i.e., foregut and midgut) is the likely site of glucose absorption, 3) the termite and its symbionts contribute specific enzymes to the digestive process, 4) …

Proteomic And Biochemical Studies Of Estrogen-Mediated Signaling And Novel Estrogen Receptor-Interacting Proteins In Breast Cancer Cells, Zhenqi Zhou

Graduate Theses and Dissertations

Estrogen plays essential roles in the growth, development, and homeostasis of a number of tissues, and can also be linked to the growth of breast cancer. The biological activities of estrogen are mediated by estrogen receptors (ERs) ERá and ERâ, and also orphan G-protein-coupled receptor 30 (GPR30). In order to identify novel proteins that are involved in ER-mediated actions of estrogen, we used mass spectrometry-based quantitative proteomic methods to systematically profile global protein expression in responses to E2 (17â-estradiol) stimulation in human breast cancer cell, and identify and characterize cellular novel proteins that are associated with ERs in breast cancer …

Design, Development, And Characterization Of Novel Antimicrobial Peptides For Pharmaceutical Applications, Yazan H. Akkam

Graduate Theses and Dissertations

Candida species are the fourth leading cause of nosocomial infection. The increased incidence of drug-resistant Candida species has emphasized the need for new antifungal drugs. Histatin 5 is a naturally occurring human salivary antifungal peptide and the first line of defense against infections of the oral cavity. This research has focused on understanding the activity of histatin 5, and subsequently designing novel peptides that may serve as models for the further development of therapeutics to treat fungal infection.

This objective has been achieved in three steps: studying the structural requirement of histatin 5 involved in antifungal activity, the identification of …

Molecular Diagnosis Of Metabolic Fast Growth Related Diseases In Broiler, Adnan Ali Khalaf Al-Rubaye

Graduate Theses and Dissertations

Pulmonary Hypertension Syndrome (PHS) and lameness are important metabolic diseases that affect rapidly growing broilers. The research reported in the first section of this dissertation focused on developing qPCR assays to identify differences in the expression levels of four candidate genes possibly associated with PHS: angiotensin II type 1 receptor (AGTR1): urotensin receptor 2D (UTS2D); serotonin receptor/transporter type 2Bn (HTR2B); and angiotensinogen cleaving enzyme (ACE). Expression levels of these candidate genes were examined in four different tissues. We established ribosomal protein S14 (RPS14) and RNA polymerase subunit 2B (RP2B) as suitable reference genes because they showed the most consistent deltaCt …

Dynamics And Model Of The Pore-Forming Protein Lysenin, Eric Krueger

Graduate Theses and Dissertations

Membrane transporters are a class of membrane proteins that function to provide a pathway across a cell membrane for the movement of ions and biomolecules. Investigations into the regulatory mechanism of these systems are hindered by their extensive preparation requirements compounded by their fragility and instability. However, lysenin, a pore-forming protein extracted from the earthworm Eisenia foetida, provided a unique opportunity to study a protein which is stable in both a soluble and membrane phase. Lysenin channels possess several important properties characteristic of ion channels without the inherent difficulties that plague investigations with biologically vital membrane transporters like voltage-gated ion …

Interaction Between Centromeric Histone H3 Variant And Shugoshin, Visarut Buranasudja

Open Access Theses

Precise and faithful segregation of chromosome segregation during mitosis depends on the ability of the cell to regulate chromosome bi-orientation on the mitotic spindle. Shugoshin (Sgo1), the protector of meiotic centromeric cohesin, is required for proper establishment of chromosome bi-orientation. Sgo1 plays a crucial role as part of a mitotic tension sensor between sister chromatids. Recently, Sgo1 has been reported to interact with histone H3 at the pericentromere region, as an important factor for tension sensing and chromosome segregation. However, the role of Sgo1 in tension sensing at centromere is still elusive. The centromere is the region of attachment of …

The Role Of Centromeric Chromatin And Kinetochore-Associated Factors In Chromosome Segregation, Wesley Williamson

Graduate Theses and Dissertations

Previous work in our lab has identified a point mutation in HTA1, one of the genes encoding histone H2A, which causes an increase-in-ploidy phenotype in Saccharomyces cerevisiae. This histone mutant strain was used to carry out a transposon insertion screen to identify suppressors of the increase-in-ploidy phenotype. This screen identified all three subunits of the Hda histone deacetylase complex, HDA1, HDA2, and HDA3. This study aims to elucidate the function of the Hda complex in chromosome segregation by exploring interactions among the members of the complex, as well as interactions between Hda complex and kinetochore components. …

Characterization And Epidemiology Of Soybean Vein Necrosis Associated Virus, Jing Zhou

Graduate Theses and Dissertations

Soybean vein necrosis disease (SVND) is widespread in major soybean-producing areas in the U.S. The typical disease symptoms exhibit as vein clearing along the main vein, which turn into chlorosis or necrosis as season progresses. Double-stranded RNA isolation and shot gun cloning of symptomatic tissues revealed the presence of a new tospovirus, provisionally named as Soybean vein necrosis associated virus (SVNaV). The presence of the virus has been confirmed in 12 states: Arkansas, Illinois, Missouri, Kansas, Tennessee, Kentucky, Mississippi, Maryland, Delaware, Virginia and New York. Symptomatic samples collected from eight states (AR, IL, MO, MS, KS, TN, MD and DE), …