Molecular Biology Commons^™

The Role Of Prolactin And Its Antagonist In Her2/Neu Tumorigenesis, Michele Scotti

All Dissertations

Purpose: To study the role of prolactin (PRL) and its antagonist, G129R, in HER2/Neu tumorigenesis. Specifically, to investigate the interaction between the oncogene HER2 and the PRL receptor (PRLR) signaling pathways for designing effective combinational therapeutics for breast cancer.
Experimental Design: The combination effects of G129R and an anti-HER2 antibody, Herceptin, were tested against HER2-overexpressing human breast cancer cell lines, T-47D and BT-474, using cell based assays and xenografts established in athymic mice. Furthermore, four different bitransgenic mouse lines co-expressing the murine version of HER2 and PRL or G129R were generated. The mammary tumor incidence, characterization of mammary gland development, …

Development Of An In Vitro Protocol To Study The Effect Of Estrogen On Osteoblast Activity, Sourik Ganguly

Morehead State Theses and Dissertations

A thesis presented to the faculty of the College of Science & Technology at Morehead State University in partial fulfillment of the requirements for the Degree of Master of Science by Sourik Ganguly on July 7, 2006.

Extracellular Superoxide Dismutase Protects Cardiomyocytes During Hyperoxia, Mervat Elsayed Ali

Theses and Dissertations in Biomedical Sciences

Reactive oxygen species are implicated in tissue damage in many cardiovascular diseases. The current study was designed to test the hypothesis that exposure to high inspired oxygen concentrations (100%) damages cardiac mitochondria and that a biogenic response is needed for cell survival. The study was done in mice with a transgene (TG) for extracellular superoxide dismutase (EC-SOD) or with the EC-SOD gene knockout (KO). Adult EC-SOD mice and their wild type littermates and EC-SOD KO mice and their wild type littermates were exposed to 100% oxygen for 6 hours and for 72 hours, and the hearts were subsequently removed under …

The Study Of Nitric Oxide Synthase Expression, Function, And Regulation In The Renal Vasculature During Postnatal Renal Development, Brian Blake Ratliff

Theses and Dissertations in Biomedical Sciences

The newborn kidney is vulnerable to vasomotor acute renal failure (ARF) from adverse perinatal events or complications of prematurity. Nitric oxide (NO) vasodilation is vitally protective in this type of ARF, but its relationship with other vasoactive factors, such as angiotensin II (AII) has not been examined. In the immature kidney, nitric oxide synthase (NOS) isoforms, specifically eNOS and nNOS, are developmentally regulated, but their specific role and regulation are unknown.

The enhanced vasodilatory role of NO in the immature kidney was hypothesized to be attributed to regulatory, expressional, and functional differences in eNOS and nNOS isoforms from the adult. …

Nanosecond Pulsed Electric Field Effects On Cell Cycle And Apoptosis, Emily H. Hall

Theses and Dissertations in Biomedical Sciences

Apoptosis, programmed cell death, is a highly regulated and complex pathway essential for embryonic development, immune-system function and maintenance of tissue homeostasis where cells induce their own cell death. Cells undergoing apoptosis exhibit a distinctive phenotype characterized by maintenance of membrane integrity, cell shrinkage, phosphatidylserine (PS) externalization at the plasma membrane, caspase protease activation, DNA fragmentation, release of cytochrome c from the mitochondrion, and membrane blebbing. An important regulatory protein in the apoptotic pathway is p53. The p53 protein functions to modulate the cell cycle by arresting cells in the G1 and G 2 phases to repair DNA damage, and/or …

Modulation Of Tgfβ-Induced Pai -1 Expression By Changes In Actin Polymerization In Human Mesangial Cells, Keyur Patel

Theses and Dissertations in Biomedical Sciences

Chronic renal diseases show increased deposition of extracellular matrix (ECM) in the glomerulus (glomerulosclerosis). Glomerulosclerosis is associated with activation of normally quiescent glomerular mesangial cells into myofibroblast-like cells. The overall objective of this study is to delineate cellular mechanism/s of myofibroblast-differentiation in disease states. In cultured mesangial cells certain characteristics of myofibroblast differentiation (α-smooth muscle actin (α-SMA) and hypertrophy) are associated with an increase in polymeric actin microfilaments (stress fibers). It is likely that other genes are also regulated in an actin cytoskeleton-dependent manner during myofibroblast differentiation. In these studies, we therefore examined the hypothesis that changes in the actin …

Use Of Cerium Oxide Nanoparticles For Protection Against Radiation-Induced Cell Death, Jimmie Colon

Electronic Theses and Dissertations

The ability of engineered cerium oxide nanoparticles to confer radioprotection was examined. Rat astrocytes were treated with cerium oxide nanoparticles to a final concentration of 10 nanomolar, irradiated with a single 10 Gy dose of ionizing radiation and cell death was evaluated by propidium iodine uptake at 24 and 48 hours after radiation insult. Treatment of rat astrocytes with nanoceria resulted in an approximate 3-fold decrease in radiation induced death. These results suggest that the nanoceria are conferring protection from radiation induced cell death. Further experiments with human cells were conducted. Human normal and tumor cells (MCF-7 and CRL8798) were …

Characterization And Evaluation Of The Immunogenizity Of Chloroplast-Derived 19-Kilodalton C-Terminal Merozoite Surface Antigen 1 (Msp1) Of Plasmodium Yoelii Yoelii, Sushamadevi Kamarajugadda

Electronic Theses and Dissertations

Malaria is a protozoan disease caused in humans by four different species of the genus Plasmodium (P. falciparum, P. vivax, P. ovale, P. malarie) and in rodents by Plasmodium yoelii yoelii. It has been reported that 1.5 to 3 million deaths occur worldwide due to malaria and the DALY (Daily affected life years) reports about 0.76% of world population affected by the disease in some of the major countries like Africa, Asia, Latin America etc., Due to the development of resistance to drugs by the parasite, there is an urgent need and prime importance for the development of an effective …

Expression Of Heterologous Proteins In Transgenic Tobacco Chloroplasts To Produce A Biopharmaceutical And Biopolymer, Andrew Leon Devine

Electronic Theses and Dissertations

The chloroplast has been demonstrated to be an ideal compartment to accumulate certain proteins or their biosynthetic products that would be harmful if they were accumulated in the cytoplasm. Hyper-expression of foreign proteins in chloroplast transgenics has accumulated up to 46% total soluble protein, this is possible due to the ~100 chloroplast genomes per chloroplast and ~100 chloroplasts per cell which can therefore, contain up to 10,000 copies of the transgene. Maternal gene inheritance of plastids in most crop plants results in natural gene containment. Chloroplast transformation also eliminates positional effects that are frequently observed with nuclear transformation and no …

Phospholipase A2mechanism Of Interfacial Activation,An Interdiscliplinary Approach, Kathleen N. Nemec

Electronic Theses and Dissertations

Phospholipase A2 (PLA2) is an enzyme that hydrolyzes the sn-2-ester bond of membrane phospholipids and liberates arachidonic acid, which is converted to eicosanoids that act as potent mediators of inflammation and allergy. As such this enzyme plays a crucial role in many homeostatic physiological and immunologic processes and disease progression. PLA2s undergo substantial increase in activity upon binding to cellular membranes. This effect of interfacial activation is well recognized, yet its structural and physical aspects are poorly understood. In this work, we have employed the interdisciplinary methods of molecular biology, biochemistry, biophysics, bioinformatics and computational biology, in order to elucidate …

Regulation Of Apoptotic Alkalinization Through Phosphorylation Of Sodium Hydrogen Exchanger Via P38 Mitogen Activated Protein Kinase, Amy Greinier

Electronic Theses and Dissertations

Regulation of intracellular pH is responsible for many cellular processes, such as metabolism, cell cycle progression, and apoptosis. Many chemotherapeutic agents work by inducing target cells to undergo apoptosis, a cell death process still poorly understood. Previous studies demonstrated that a rise in intracellular pH activated apoptotic proteins leading to cytochrome C release. This "apoptotic alkalinization" occurred upon activation of the plasma membrane protein, sodium hydrogen exchanger-1 (NHE1), whose activity is regulated by the stress kinase p38 MAPK. In previous studies, upon cytokine withdrawal from cytokine-dependent lymphocytes induced the activity of the p38 MAP kinase which then phosphorylated the C-terminus …

Expression And Characterization Of Mycobacterium Paratuberculosis 19kda With Posttranslational Modification, Mitra Safavi-Khasraghi

Electronic Theses and Dissertations

Despite the fact that E. coli supports limited posttranslational modification, this bacterium has been universally used as the expression system of choice. Expression of modified proteins in E. coli may lead to expression of recombinant proteins that lack essential immunomodulatory or catalytic components essentials for infectious processes. Previously in our laboratory, pMptb#28 plasmid containing a 4.8 kb insert from M. paratuberculosis has been identified which expressed 16 kDa recombinant protein in E. coli and 19 kDa recombinant protein in Mycobacterium smegmatis. The objective of this study is to identify the ORF sequence, investigate possible posttranslational modification and characterize the protein …

Bioinformatic Analysis Of Solanaceae Chloroplast Genomes And Characterization Of An Arabidopsis Protein Disulfide Isomerase In Transgenic Tobacco Chloroplasts, Justin James Grevich

Electronic Theses and Dissertations

Throughout history, traditional plant breeding has been used to provide resistance to pests, disease and other forms of environmental stress, as well as to increase yield and improve upon quality and processing attributes. Over the last decade, the advancement in sequencing technology and bioinformatic analysis has unleashed a wealth of knowledge about chloroplast genetic organization and evolution. The lack of complete plastid genome sequences is one of the major limitations in advancing plastid genetic engineering to other useful crops. This is due to the fact that plastid genome sequences are essential for the identification of endogenous regulatory sequences and optimal …

Studies On The Novel Function Of Amyloid Precursor Protein In Glial Differentiation Of Neural Stem Cells, Young-Don Kwak

Electronic Theses and Dissertations

Although amyloid β (Aβ) deposition has been a hallmark of Alzheimer’s disease (AD), the physiological function of amyloid precursor protein (APP) is not clear. Our results suggested that high concentration of APP induces glial differentiation while physiological level of APP promotes migration and differentiation of neural stem cell (HNSC). HNSCs were mainly differentiated into astrocytes when they are transplanted into APP transgenic mouse brain or treated with a high concentration of secreted-type APP (sAPP) in culture. Staurosporine (STS) induced a distinctive astrocytic morphology in NT-2/D1 neural progenitor cells with expressions of APP and astrocyte-specific markers, glial fibrillary acidic protein (GFAP), …

Isolation And Functional Mapping Of Human T-Cell Leukemia Virus Type 1 Tax Oncoprotein Dna-Damage Complexes, Sarah Saionz Durkin

Theses and Dissertations in Biomedical Sciences

Human T-cell Leukemia Virus Type 1 (HTLV-1) is a transforming retrovirus which causes Adult T-cell Leukemia (ATL) and HTLV-1 Associated Myelopathy/Tropical Spastic Paraparesis (HAM/TSP). Cellular transformation can be caused by a single viral trans-activating protein, Tax. Tax may contribute to transformation through interaction with components of the DNA damage response pathway, promoting cellular genomic instability. We examined cellular Tax complexes in an effort to elucidate potential protein-protein interactions that can model the Tax-induced molecular events.

We also investigated the role of post-translational modification in regulating Tax function. We employed a direct physical analysis of Tax complexes isolated from mammalian …

The Use Of Proteomic Technologies To Identify Serum Glycoproteins For The Early Detection Of Liver And Prostate Cancers, Elizabeth Ellen Schwegler

Theses and Dissertations in Biomedical Sciences

The application of proteomic technologies to identify serum glycoproteins is an emerging technique to identify new biomarkers indicative of disease severity. Many of these newly evolving protein-profiling methodologies have evolved from previous global protein expression profiling studies such as those involving SELDI-TOF-MS technologies. Though the SELDI approach could distinguish disease from normal by utilizing protein patterns as shown herein with the HCC study of chapter II, it was unable to offer sequence information on the selected peaks, and did not have the ability to analyze the entire dynamic range of the serum/plasma proteome. To address these deficiencies, new strategies that …

Molecular Epidemiology Of Antibiotic Resistant Salmonella Enterica Strains From Different Animal And Human Sources In Ireland, David George Lee

Theses

In this study, 176 isolates of Salmonella were collected from different laboratories in Ireland. All isolates were confirmed to be of the genus Salmonella by performing bacteriological and biochemical tests. Briefly, all isolates were cultured onto XLD agar after which pure cultures were verified and stocked at this institute. Definitive identification was carried out using the API 20-E system and a separate urease test was also performed on all isolates. After confirmation of the genus, all isolates were prepared for antibiotic susceptibility testing and r-types were determined using sixteen antibiotics. Depending on the antibiotic resistance profiles, isolates were selected for …

A Molecular Technique For Specific Identification Of Western Atlantic Ocean Scombrids And An Analysis Of A Larval Scombrid Assemblage Off The Kona Coast Of Hawaii, Melissa A. Paine

Dissertations, Theses, and Masters Projects

No abstract provided.