Life Sciences Commons^™

Effects Of Metal Protoporphyrins On Burn Wound Conversion, Katie Braun

Theses and Dissertations

A murine model was utilized to test the influence of heavy metal protoporphyrins on burn conversion, heme oxygenase – 1 (HO-1) expression, and inflammation. Heavy metal protoporphyrins, such as cobalt protoporphyrin (Co PP) and tin protoporphyrin (Sn PP), were used to influence the heme oxygenase activity. The effects of these heavy metal protoporphyrins on burn wound conversion were examined using a burn comb model in rats. In addition to assessing the extent of conversion, HO-1 expression and parameters of inflammation were also examined in the area of injury (interspace region) subject to conversion. These studies demonstrate proof in principal that …

Effects Of Light Exposure On The Release Of Oxygen From Hemoglobin In A Red Blood Cell Suspension, Tanikka Toler

Theses and Dissertations

The main function of the cardiovascular system is to deliver a sufficient quantity of oxygenated blood to the tissues, cells, and organs of the body in order to provide the cells with essential nutrients for metabolism and for the removal of waste products. All cells require and utilize oxygen. Oxygen is transported to various cells and tissues via red blood cells flowing through the microcirculation of an organism. Measurement of oxygen transport in the microcirculation has shown that about ten times more oxygen appears to leave the blood of arterioles than can be accounted for by diffusion. One possibility to …

Molecular Mechanisms Involved In Oligodendrocyte Development, Rochelle Coelho

Theses and Dissertations

Multiple sclerosis (MS) is a chronic disease of the central nervous system (CNS) characterized by demyelination and loss of oligodendrocytes (OLGs), the CNS myelin-producing cells. Thus, understanding the mechanisms that control OLG development can provide valuable knowledge regarding remyelination therapies for MS. This disease is thought to result from an autoimmune attack towards myelin. FTY720, an immunomodulator under evaluation for MS treatment is a sphingosine-1-phosphate (S1P) analog. We found before that S1P plays a crucial role in the NT-3-mediated survival of OLGs, an observation that led us to investigate whether FTY720 could have any effect on these cells. Our studies …

Regulation Of Mdm2 Mediated Nfκb2 Pathway In Human Lung Cancer, Lathika Mohanraj

Theses and Dissertations

Overexpression of oncoprotein MDM2 and mutations of tumor suppressor p53 are frequently observed in human cancers. The NFκB pathway is one of the deregulated pathways in oncogenesis. The overall goal of the project was to study the regulation of NFκB pathway by MDM2 in lung cancer. Our first effort was to determine the frequency of MDM2 overexpression in human lung tumor samples and to identify co-occurring abnormal gene expression by studying the levels of MDM2 and members of NFκB pathway with respect to p53 status. Higher than normal levels of MDM2 were found in approximately 30% of the cancer samples …

Co-Administration Of Sildenafil Potentiates Doxorubicin-Induced Apoptosis In Prostate Cancer: The Role Of Nf-Kappab, Sarah Hassanieh

Theses and Dissertations

Our recent studies have shown that that erectile dysfunction (ED) drugs including Sildenafil (Viagra), Vardenafil (Levitra) and Tadalafil (Cialis) enhance killing of several types of cancer cells by anticancer drug, Doxorubicin (DOX). We observed increased cell death by apoptosis in response to the combined treatment with ED drugs and DOX. However, the mechanism of such enhancement of cell death by combined treatment of ED drugs and DOX is not fully understood. Nuclear factor-κB (NF-κB) is an oxidant-sensitive transcription factor that plays a critical role in the immediate-early activation of a multitude of genes that have been documented to play critical …

Does Light Control Algal Abundance In Large River Systems?, Amy Macdonald

Theses and Dissertations

A limited amount of research has been done to investigate the factors influencing algal abundance in large river systems. This study examines light as the primary factor that controls algal abundance in the Upper Mississippi, Missouri and Ohio Rivers. Data were collected for 2004 in conjunction with the Environmental Monitoring Assessment Program- Great River Ecosystems EMAP-GRE project using EPA approved methods. Chlorophyll a concentrations were 34.6 µg•L-1 in the Upper Mississippi, 19.8 µg•L-1 in Missouri River and 9 µg•L-1 in the Ohio River for 2004. Chlorophyll a concentrations were significantly different among the three rivers (p<0.0001) but not between years. Inter-river variation could be loosely correlated with light availability: mean Average Irradiance Dosages, which consider factors that affect light climate (depth, transparency, velocity, surface irradiance), by river corresponded with mean chlorophyll a levels by river. Intra-river variation seemed to be due to both the influence of light and nutrients.

Crosstalk Between Mdm2 And Akt Signaling Pathway In Oncogenesis., Mahesh Ramamoorthy

Theses and Dissertations

MDM2, the human homologue of the Mouse Double Minute 2 gene product, has been shown to be over-expressed in many cancers and to induce tumorigenesis. The role of MDM2 in oncogenesis was thought to be p53 dependent. However recent years have shown MDM2 to be a key player in a complex network of interactions that affect cell cycle, apoptosis, and tumorigenesis in a p53 independent manner. Here we report a novel p53 independent role for the multidimensional protein MDM2; its ability to induce phosphorylation of Akt at serine 473 residue. Transient and stable over-expression of MDM2 in cultured cell lines …

Ceramide Kinase And Ceramide-1-Phosphate, Dayanjan Wijesinghe

Theses and Dissertations

Ceramide-1-phosphate (C1P) is a bioactive lipid that has been implicated in many biological processes. Our laboratory has conclusively demonstrated its role in inflammation via activation of cPLA2α. The only known enzyme to date responsible for direct synthesis of C1P is ceramide kinase. Very little was known about this enzyme in terms of its enzyme kinetics and substrate specificity. As CERK is an enzyme that acts on membrane lipids, its kinetics cannot be studied using standard bulk dilutions methods. Thus we developed a surface dilution approach using Triton X 100 mixed micelles for studying the kinetics of CERK. We discovered that …

Avian Dispersal Of The Actinomycete Frankia Across A Barrier Island Landscape, Spencer Bissett

Theses and Dissertations

In the nutrient-poor soils characteristic of coastal environments, symbiotic association with the nitrogen-fixing root endosymbiont Frankia is essential to establishment and survival of the woody shrub Morella cerifera. Nutrient deficiency quickly becomes severe unless seedlings are infected by Frankia soon after germination. However, the means of arrival of Frankia prior to shrub establishment has not been determined. Using sterilized lab-grown M. cerifera seedlings and fecal samples collected from passerine birds on the Eastern Shore of Virginia, viability of avian dispersal of the bacteria was tested. Although passerine fecal samples did produce nodules on some sterilized M. cerifera seedlings, these experimental …

Application Of Shortest-Path Network Analysis To Identify Genes That Modulate Longevity In Saccharomyces Cerevisiae, Jr Managbanag

Theses and Dissertations

Shortest-path network analysis was employed to identify novel genes that modulate longevity in the baker’s yeast Saccharomyces cerevisiae. Based upon a set of previously reported genes associated with increased life span, a shortest path network algorithm was applied to a pre-existing protein-protein interaction dataset in order to construct a shortest-path longevity network. To validate this network, the replicative aging potential of 88 single gene deletion strains corresponding to predicted components of the shortest path longevity network was determined. The 88 single-gene deletion strains identified by a network approach are significantly enriched for mutation conferring both increased and decreased replicative life …

The Role Of Microrna-155 And Microrna-146a As Putative Oncomirs In The Tumor Progression Of Prostate Cancer, Jennifer Hoyt

Theses and Dissertations

Prostate cancer is the most common cancer occurring in males. The identification of novel microRNAs (miRs) that contribute to tumor progression represents prospective treatment targets. miRs are small non-coding RNAs important in gene regulation with specific tissue expression patterns. Each miR is thought to affect the expression of hundreds of different RNA targets. Two putative oncomiRs, miR-155 and miR-146a, were shown to be differentially expressed in the human derived, prostate cell sublines M12 and F6. Quantification of endogenous miR expression showed high levels in the metastatic M12 cell line versus low in its weakly tumorigenic F6 variant. The restoration of …

The Response Of Hn4 Cells To Porphyromonas Gingivalis Dna, Cheyanne Warren

Theses and Dissertations

Periodontal disease is one of the most common human diseases. Bacteria trigger the onset and progression of the disease and among them Porphyromonas gingivalis has been demonstrated to be a major etiologic agent. Although the interaction of the bacterium with the host is of major importance for the understanding of the disease mechanisms, both the host as well as the pathogen components involved in the interaction remain poorly understood. One of the bacterial components capable of eliciting a host response is unmethylated CpG DNA motifs found in bacteria. Thus, the first aim was to determine the response of oral epithelial …

Plumage Ornamentation As An Indicator Of Female Age And An Influence In Male Mate Choice In Protonotaria Citrea, The Prothonotary Warbler In Virginia., Terry Smith

Theses and Dissertations

Flamboyant plumage and ornamentation is common and well-known in male birds; it serves as a sexual display to attract potential mates. While flamboyant plumage is less common and usually more subtle in female birds, it does occur in some species such as Dark-Eyed Juncos (Junco hyemalis) and Prothonotary Warblers (Protonotaria citrea). Prothonotary Warblers display relatively subtle sexual dimorphism. This study examines variations in tail spot patterns in Prothonotary Warblers and relates those variations to age in females. Females with fewer than six spots tend to be two years old or younger; females with six spots or more tend to be …

Intravenous Administration Of Perfluorocarbon Emulsions As A Non-Recompression Therapy For Decompression Sickness, Cameron Smith

Theses and Dissertations

Decompression sickness (DCS) results from a sudden decrease in ambient pressure leading to super-saturation of tissues with inert gas and subsequent bubble formation within both tissues and blood. Perfluorocarbons (PFC) are able to dissolve vast amounts of non-polar gases. The administration of intravenous (I.V.) PFC emulsions reduce both morbidity and mortality of DCS, but the mechanism of this protective effect has not yet been demonstrated. Juvenile Dorper cross sheep between 16 and 24 kg (n=31) were anaesthetized and instrumented for physiological monitoring, the administration of I.V. fluids and sampling of arterial and mixed venous blood. Animals were placed in a …

Effect Of Hemoglobin-Based Oxygen Carriers On Arterial Pressure And Vasoactivity In The Rat Mesentery, Michael Kim

Theses and Dissertations

Hemoglobin-based oxygen carriers provide a promising future as an alternative to human blood transfusions. Hemoglobin-based oxygen carriers, HBOCs, provide a lowcost, easy to maintain, and safe solution. They require no refrigeration and are universally compatible, and the required transfusion volume is less than that of a normal transfusion. HBOCs have been known to have adverse side effects such as renal toxicity, gastrointestinal dismotility, and hypertension. Many of these problems stem from the lack of a membrane, which protects the hemoglobin from dissociating and extravasating into the blood vessel wall. Extracellular hemoglobin, like that found in HBOCs, has a greater affinity …

The Effect Of Nacl On Akinete Differentiation In The Cyanobacterium Nostoc Punctiforme, Jonathan Heekin

Theses and Dissertations

Nostoc punctiforme is a nitrogen-fixing, symbiotic/free-living cyanobacterium. There has been a great deal of research conducted on the genomic nature of N. punctiforme as it pertains to its ecologically important role in the nitrogen cycle in varied environments around the world. My study concentrated on the dormant cell type known as the akinete. Increasing concentrations of NaCl were used to follow the growth phases from germination to akinete formation (lag phase-logarithmic growth phase-stationary phase). I found that increased salt concentrations caused N. punctiforme to form akinetes faster when compared to the control. Germination rates were not greatly increased or shortened …

Effects Of Hemoglobin-Based Oxygen Carriers On The Vasoactivity Of The Spinotrapezius Muscle Of The Rat, Pete Meliagros

Theses and Dissertations

Hemoglobin-based oxygen carriers (HBOCs) offer a safe, more plentiful and long term alternate to blood banks. However, they have been found to increase blood pressure which can be attributed to an increase in total peripheral resistance (TPR). Lumenal nitric oxide (NO) scavenging by these HBOCs seems to be responsible for this hypertensive effect. In addition, it is believed that hemoglobin (Hb) tetramers and dimers may extravasate and consume additional nitric oxide in the perivascular and interstitial space. The purpose of the present study was to elucidate the role of NO scavenging and to confirm extravasation as a contributor to HBOC …

The Ecology Of Fear: Oviposition And Colonization In Aquatic Systems, Leeanna Pletcher

Theses and Dissertations

Amphibians and aquatic invertebrates have complex life histories that link aquatic and terrestrial food webs. It has been suggested that amphibian reproduction is an important source of carbon to some aquatic systems. This process of energy flow may be shaped by shifts in habitat selection in response to predators. We hypothesized that predators decrease colonization and oviposition of prey, reducing active inputs. Thus predation risk is expected to shift the relative amounts of active and passive subsidies. We manipulated the presence of fish predators in aquatic mesocosms. Results suggest hylid treefrog eggs and hydrophilid beetles were less abundant in predator …

Effects Of Predation Risk, Density And Disease On Energy Efficiency In A Larval Anuran, Sarah Crane

Theses and Dissertations

Predation, density and disease affect behavior, morphology and growth. There is a lack of information on how these changes relate to efficiency of energy transfer in anuran larvae, although previous studies suggest that predation should decrease and competition should increase efficiency. Using a 2 x 2 factorial design, we manipulated predation presence and larval density to test how predation risk and density affect energy efficiency. During the experiment, approximately half of the tadpoles were infected by an unknown disease. Neither predation risk nor density affected assimilation or growth efficiency, despite changes in growth and development. Disease, however, decreased gut length …

Biological Functions Of The Novel Lysophosphatidic Acid (Lpa) Receptor, Lpa4/P2y9/Gpr23, Peilun Lee

Theses and Dissertations

Lysophosphatidic acid (LPA), a naturally occurring phospholipid present in serum and malignant effusions, elicits diverse biological functions through binding and activating specific cell surface G-protein coupled receptors. In addition to the conventional LPA1/Edg2, LPA2/Edg4 and LPA3/Edg7 receptors of the endothelial differentiation gene (Edg) family, LPA4/p2y9/GPR23 of the purinergic receptor family and the related LPA5/GPR92 and LPA6/p2y5 have been identified as novel LPA receptors. These newly identified LPA receptors are structurally distant from the Edg LPA1-3 receptors and couple to Gq, G12/13 and probably Gs subunits. However, the roles of the LPA4-6 receptors in LPA signal transduction and physiology are poorly …

The Ecology Of Fear: Colonization And Oviposition In Aquatic Systems [Poster], Leeanna Pletcher, Johanna Kraus, James R. Vonesh

Biology Presentations

Introduction

Amphibians and aquatic invertebrates have complex life histories that link aquatic and terrestrial food webs. It has been suggested that amphibian reproduction is an important source of carbon to some aquatic systems. Movement of organisms across the aquatic-terrestrial habitat boundary can represent important subsidies to the receiving habitat. Subsidies are organisms, nutrients, or detritus that cross habitat boundaries and are consumed, and these allochthonous inputs can affect food web structure. Predators can alter subsidies by consuming organisms that would otherwise move across habitat boundaries. Predator induced shifts in habitat selection are a well known non-lethal effect in aquatic systems. …

Nadph Oxidase As A Therapeutic Target In Alzheimer's Disease, Michelle L. Block

Anatomy and Neurobiology Publications

At present, available treatments for Alzheimer's disease (AD) are largely unable to halt disease progression. Microglia, the resident macrophages in the brain, are strongly implicated in the pathology and progressively degenerative nature of AD. Specifically, microglia are activated in response to both β amyloid (Aβ) and neuronal damage, and can become a chronic source of neurotoxic cytokines and reactive oxygen species (ROS). NADPH oxidase is a multi-subunit enzyme complex responsible for the production of both extracellular and intracellular ROS by microglia. Importantly, NADPH oxidase expression is upregulated in AD and is an essential component of microglia-mediated Aβ neurotoxicity. Activation of …

A Synaptic Nidogen: Developmental Regulation And Role Of Nidogen-2 At The Neuromuscular Junction, Michael A. Fox, Matthew Sp Ho, Neil Smyth, Joshua R. Sanes

Anatomy and Neurobiology Publications

Background

The skeletal neuromuscular junction is a useful model for elucidating mechanisms that regulate synaptogenesis. Developmentally important intercellular interactions at the neuromuscular junction are mediated by the synaptic portion of a basal lamina that completely ensheaths each muscle fiber. Basal laminas in general are composed of four main types of glycosylated proteins: laminins, collagens IV, heparan sulfate proteoglycans and nidogens (entactins). The portion of the muscle fiber basal lamina that passes between the motor nerve terminal and postsynaptic membrane has been shown to bear distinct isoforms of the first three of these. For laminins and collagens IV, the proteins are …

Arabidopsis Mrna Polyadenylation Machinery: Comprehensive Analysis Of Protein-Protein Interactions And Gene Expression Profiling, Arthur G. Hunt, Ruqiang Xu, Balasubrahmanyam Addepalli, Suryadevara Rao, Kevin P. Forbes, Lisa R. Meeks, Denghui Xing, Min Mo, Hongwei Zhao, Amrita Bandyopadhyay, Lavanya Dampanaboina, Amanda Marion, Carol Von Lanken, Qingshun Quinn Li

Anatomy and Neurobiology Publications

Background

The polyadenylation of mRNA is one of the critical processing steps during expression of almost all eukaryotic genes. It is tightly integrated with transcription, particularly its termination, as well as other RNA processing events, i.e. capping and splicing. The poly(A) tail protects the mRNA from unregulated degradation, and it is required for nuclear export and translation initiation. In recent years, it has been demonstrated that the polyadenylation process is also involved in the regulation of gene expression. The polyadenylation process requires two components, the cis-elements on the mRNA and a group of protein factors that recognize the cis …

Shortest-Path Network Analysis Is A Useful Approach Toward Identifying Genetic Determinants Of Longevity, J. R. Managbanag, Tarynn M. Witten, Danail Bonchev, Lindsay A. Fox, Mitsuhiro Tsuchiya, Brian K. Kennedy, Matt Kaeberlein

Study of Biological Complexity Publications

Background

Identification of genes that modulate longevity is a major focus of aging-related research and an area of intense public interest. In addition to facilitating an improved understanding of the basic mechanisms of aging, such genes represent potential targets for therapeutic intervention in multiple age-associated diseases, including cancer, heart disease, diabetes, and neurodegenerative disorders. To date, however, targeted efforts at identifying longevity-associated genes have been limited by a lack of predictive power, and useful algorithms for candidate gene-identification have also been lacking.

Methodology/Principal Findings

We have utilized a shortest-path network analysis to identify novel genes that modulate longevity in Saccharomyces …

Protocol: Streamline Cloning Of Genes Into Binary Vectors In Agrobacterium Via The Gateway® Topo Vector System, Ruqiang Xu, Qingshun Quinn Li

Anatomy and Neurobiology Publications

Background

In plant functional genomic studies, gene cloning into binary vectors for plant transformation is a routine procedure. Traditionally, gene cloning has relied on restriction enzyme digestion and ligation. In recent years, however, Gateway^® cloning technology (Invitrogen Co.) has developed a fast and reliable alternative cloning methodology which uses a phage recombination strategy. While many Gateway- compatible vectors are available, we frequently encounter problems in which antibiotic resistance genes for bacterial selection are the same between recombinant vectors. Under these conditions, it is difficult, if not sometimes impossible, to use antibiotic resistance in selecting the desired transformants. We have, …

Augmentation Of T Cell Expansion For Adoptive Immunotherapy By Alternate Gamma Chain Cytokines And By Gemcitabine Mediated Inhibition Of Myeloid Derived Suppressor Cells, Hanh Le

Theses and Dissertations

Successful treatment of cancer with adoptive immunotherapy (AIT) is dependent on the ability to produce large numbers of tumor-specific, functional T cells. The purpose of this thesis is to explore ways in which T cell expansion could be augmented. We have focused on exploring alternate gamma chain cytokines as stimulators of T cell proliferation and differentiation in addition to investigating the potential usefulness of gemcitabine (GEM) in abrogating the immunosuppressive effects of myeloid derived suppressor cells (MDSCs). B16 melanoma sensitized draining lymph node cells that have been activated in vitro with bryostatin-1 and ionomycin (B/I) were expanded in either IL-7/15 …

The Effect Of Cell Type On The Efficacy Of Cmv Antiviral Drugs, Benjamin Meza

Theses and Dissertations

Until recently, all in vitro drug susceptibility assays of cytomegalovirus (CMV) were performed in clinically irrelevant fibroblast cells. This study sought to test if drug susceptibility was affected by cell type. MRC-5 embryonic lung fibroblasts and ARPE-19 retinal pigmented epithelial cells were infected with BADrUL131-Y4 epithelial/fibroblast tropic virus under serial concentrations of ganciclovir (GCV) or maribavir (MBV). Virus was quantified using plaque reduction, GFP fluorescence, and yield reduction. Both drugs performed less efficiently in ARPE-19 cells. A cell type effect was observed for both plaque reduction and yield reduction assays with implications for the treatment of CMV retinitis as well …

Role Of The Small Terminase Subunit Encoded By Staphylococcus Aureus Pathogenicity Island Sapi1 In Formation Of Sapi1 Transducing Particles, Nicholas Paul Olivarez

Theses and Dissertations

Staphylococcus aureus pathogenicity island SaPI1 is a genomic element that is mobilized and transduced at high frequency by helper phage 80α. SaPI1 encodes a small terminase protein that belongs to the phage small terminase subunit family. The presence of SaPI1-encoded small terminase suggests that it plays a role in SaPI1-specific packaging into transducing particles by complexing with the 80α large terminase subunit and redirecting recognition to a pac site on SaPI1 DNA from 80α DNA. The effects of deleting the small terminase genes in SaPI1 and in a prophage copy of 80α are consistent with this hypothesis. Induction of the …

Inflammation-Associated Gene Regulation In Primary Astrocytes, Glial Tumors And Cellular Differentiation, Katarzyna Marta Wilczynska

Theses and Dissertations

This dissertation elucidates several independent molecular mechanisms that function in astrocytes and glial tumor cells, and suggest that developmental and inflammatory signals may contribute to the development of brain tumors. First, we analyzed the mechanism of TIMP-1 activation in astrocytes and glioblastoma cells. TIMP-1 expression is activated by IL-1, which is the major neuroinflammatory cytokine, via simultaneous activation of IKK/NF-kB and MEK3/6/p38/ATF-2 pathways in primary human astrocytes. In contrast to astrocytes, TIMP-1 is expressed at lower levels in glioblastomas, and is not regulated by IL-1 due to either dysfunctional IKK/NF-kB or MEK3/6/p38/ATF-2 activation. Thus, we propose a novel mechanism of …