Life Sciences Commons^™

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 …

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 …

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, …

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. …

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 …

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 …

Mechanical Abrasion And Electroporation In The Transformation Of Intact Pleurotus Ostreatus Hyphæ And Nutrient-Dependant Resistance To Hygromycin B, Alexander Cordesman

Theses and Dissertations

ABSTRACT 1: Abrasive treatment and subsequent electroporation of the basidiomycete Pleurotus ostreatus in a hypertonic buffer was investigated as a potential method of transforming of intact hyphæ. Mycelia, which are not capable of being transformed via electroporation due to interference by the cell wall, were abraded in an attempt to mechanically degrade the cell wall prior to electroporation. An electroporation field strength of 12,500 V/cm for 500 μs to 1.25 ms was found to be optimal based upon mortality effects. A 32 μm carborundum abrasive was initially evaluated but was quickly found to be inappropriately large so a corundum abrasive …

Role Of Vegf-C In Proliferation And Migration In A Cancer Model, Emily Marie Benke

Theses and Dissertations

Head and neck cancer ranks high among the most common cancers world wide. In addition, there is a high recurrence rate, as well as a high prevalence of loco-regional tumor spread. Among many factors contributing to metastasis is vascular endothelial cell growth factor C. VEGF-C is primarily an inducer of new lymph vessel formation, typically during embryogenesis; however, some advanced cancers show a significant increase in VEGF-C expression, suggesting a role in metastasis. In the current study, the effects of VEGF-C expression were tested in HN12 cells, which are highly metastatic and known to express high levels of the chemokine …

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 …

Camk-Ii Promotes Beta-Catenin-Dependent Transcription By Binding Flightless-I, Jamie Mcleod

Theses and Dissertations

Transient intracellular elevations of Ca2+ are common signaling mechanisms used to allosterically regulate proteins. One potential target of Ca2+ is Ca2+/calmodulin dependent protein kinase type II (CaMK-II). CaMK-II is a multi-functional protein kinase known to influence cellular pathways such as cell motility and cell cycle progression. Within the cell cycle, CaMK-II promotes the expression of the regulator protein cyclin D1, which is necessary for cell cycle progression. To further understand CaMK-II’s role in cyclin D1 expression, the binding partners of cytosolic CaMK-II were studied using mass spectrometry. Several proteins were identified including β actin, β tubulins, tropomodulin- 3 and Flightless-I. …

Fibronectin-Dependent Activation Of Camk-Ii Promotes Focal Adhesion Turnover By Inducing Tyrosine Dephosphorylation Of Fak And Paxillin, Charles Easley Iv

Theses and Dissertations

Transient elevations in Ca²⁺ have previously been shown to promote focal adhesion disassembly and cell motility. Yet the targets of these Ca²⁺ transients have not been fully examined. In this study, we demonstrate that CaMK-II, a Ca²⁺/calmodulin dependent protein kinase, is activated in response to β1 integrin engagement with fibronectin to influence fibroblast adhesion and motility. We also show that CaMK-II is dynamically localized to the cell surface using Total Internal Reflection Fluorescence microscopy (TIRFm) and that inhibition of CaMK-II with two mechanistically distinct, membrane permeant inhibitors accelerates spreading on fibronectin, enlarges paxillin-containing focal adhesions and …

Regulation Of Platelet-Activating Factor Acetylhydrolase By Oxidized Phospholipids And Proinflammatory Cytokines, Muralikrishna Mukkamala

Theses and Dissertations

Platelet-Activating Factor Acetylhydrolase (PAFAH) is a monocyte-derived phospholipase A2 that catalyzes the hydrolysis of platelet-activating factor (PAF) and has been implicated in atherosclerosis. Although PAF and other proinflammatory stimuli are postulated to induce the enzyme, mechanisms controlling PAFAH expression are largely unknown at present. We have shown that PAFAH induction in monocytes is increased in response to oxidized phospholipids. The PAFAH 5' flanking region has at least 10 putative Stat elements, and IL-6 has been shown to be downstream from the prostaglandin receptor, EP2, which has been shown to bind oxidized phospholipids, prompting the hypothesis that Stat proteins might regulate …

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 …

The Role Of Secreted Phosphoprotein-24 In Osteoblast Differentiation And Matrix Mineralization, Samuel Ramage

Theses and Dissertations

Secreted Phosphoprotein-24 (Spp24) was initially isolated and characterized as a component of bovine cortical bone matrix. Subsequent characterization has shown it is multiply phosphorylated and homologous to cystatin and TGF-β receptor type II. Spp24 is a minor component of the serum fetuin mineral complex that binds calcium-phosphate minerals and prevents their deposition. The TGF-β receptor homology domain binds BMP-2 weakly in vitro and enhances BMP-2’s osteogenic effects in vivo. The ability of Spp24 to affect BMP activity suggests an important role for Spp24 as a native, bioactive componentof bone that regulates bone development.

Spp24 was highly up-regulated in rat cortical …

Stream Ambient Noise, Spectrum And Propagation Of Sounds In The Goby Padogobius Martensii: Sound Pressure And Particle Velocity, Marco Lugli, Michael L. Fine

Biology Publications

The most sensitive hearing and peak frequencies of courtship calls of the stream goby, Padogobius martensii, fall within a quiet window at around 100Hz in the ambient noise spectrum. Acoustic pressure was previously measured although Padogobius likely responds to particle motion. In this study a combination pressure (p) and particle velocity (u) detector was utilized to describe ambient noise of the habitat, the characteristics of the goby’s sounds and their attenuation with distance. The ambient noise (AN) spectrum is generally similar for p and u (including the quiet window at noisy locations), although the energy distribution of uspectrum is …

The Legacy Of Harvest And Fire On Ecosystem Carbon Storage In A North Temperate Forest, Christopher M. Gough

Biology Publications

Forest harvesting and wildfire were widespread in the upper Great Lakes region of North America during the early 20th century. We examined how long this legacy of disturbance constrains forest carbon (C) storage rates by quantifying C pools and fluxes after harvest and fire in a mixed deciduous forest chronosequence in northern lower Michigan, USA. Study plots ranged in age from 6 to 68 yrs and were created following experimental clear-cut harvesting and fire disturbance. Annual C storage was estimated biometrically from measurements of wood, leaf, fine root, and woody debris mass, mass losses to herbivory, soil carbon content, and …

Role Of Rok And Pkc In Permeabilized Rabbit Femoral Artery, Lyndsay Jacquelyn Clelland

Theses and Dissertations

Discoveries made with KCl-induced contractions have elucidated the more complex reactions involved in GPCRs signaling; once the mechanisms of smooth muscle Ca2+ sensitization and desensitization are fully understood, then the development of advanced treatments for vascular disorders such as hypertension, cerebral and coronary vasospasm, and vascular hyporeactivity following hemorrhagic shock may be possible. Studies have shown that KCl-induced contractions induce Ca2+-sensitization. Therefore, we tested the hypothesis that KCl induced Ca2+-sensitization is due to ROK activation by the increase in [Ca2+]i. To test this hypothesis, rabbit femoral arteries were permeabilized with 20µg/ml α-toxin and 1% Triton X-100 and subjected to different …

Corticular Photosynthetic Dynamics For A Coastal Evergreen Shrub: Myrica Cerifera, Jaclyn K. Vick

Theses and Dissertations

I quantified seasonal variations in corticular photosynthesis in 1st through 5th order branches of Myrica cerifera L. (Myricaceae) in order to determine whether corticular photosynthesis contributes to whole plant carbon gain by reducing respirational CO2 loss. Maximum % refixation was 110 ± 39 % of CO2 efflux in the dark (Rd) in 1st order branches during winter, minimum was 18 ± 3 % in 5th order branches during summer. Variations in % refixation paralleled changes in photosynthetically active radiation (PAR). As light attenuated with increasing branch order % refixation decreased. Increased PAR in the winter due to a more sparse …

The Role Of Sphingosine Kinase 2 In Cell Growth And Apoptosis, Heidi M. Sankala

Theses and Dissertations

Two isoforms of sphingosine kinase (SphK) catalyze the formation of sphingosine-1-phosphate (SIP). Whereas, SphKl stimulates cell growth and survival, it was found that when overexpressed in mouse NIH 3T3 fibroblasts SphK2 enhances caspase-dependent apoptosis in response to serum deprivation, independently of S1P receptors. Sequence analysis revealed that SphK2 contains a 9 amino acid motif similar to that present in BH3-only proteins. Studies showed that the BH3-only domain, catalytic activity, endoplasmic reticulum (ER) stress, and uptake of calcium by the mitochondria may all contribute to the apoptotic effects of overexpressed SphK2 in NIH 3T3 cells. Further studies in human carcinoma cells …