Life Sciences Commons^™

Quantitative Analysis Of Glycinergic Neurons Including Ia Inhibitory Interneurons In The Ventral Spinal Cord Using A Bac-Glyt2-Egfp Transgenic Mouse Model, Palak Rajeshkumar Painter

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We are interested in understanding the development of reciprocal inhibition, an important sensory-motor circuit that prevents co-contraction of antagonist muscles during locomotion. This effect is mediated by a subset of glycinergic neurons termed Ia inhibitory interneurons (IaINs). Transgenic mouse models are useful in helping to identify subsets of neurons in developing animals. In this thesis we first wanted to test the BAC-GlyT2-eGFP mice model by measuring GFP expression overlap in neurons with glycine in late-stage embryos (E17.5) and early postnatal animals (P0/P1). Our results suggest more than 98% of glycinergic neurons at these stages also express GFP in GlyT2-EGFP transgenic …

House Dust Mite Induced Gene Expression And Cytokine Secretion By Human Dermal Fibroblasts, Jananie Rockwood

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House dust mites are a group of mites that are commonly found in homes worldwide. The three most commonly found species of house dust mites in temperate environments in the United States are Dermatophagoides farinae, Dermatophagoides pteronyssinus and Euroglyphus maynei. House dust mite allergens are strongly associated with asthma, perennial rhinitis and atopic dermatitis. Whole dust mite extracts contain both allergen and non-allergen molecules that can influence cell functions in the skin, mucous membranes of the nose and eyes, and epithelial linings of the lungs. Extracts from the three different house dust mite species contain different molecules that induce different …

Characterization Of Hiv-1 Rev Mutants, Jennifer Lynn Chang

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The HIV Rev protein has the ability to bind tubulin heterodimers and depolymerize microtubules (MTs) in vitro (Watts et al. 2000). These interactions may account for MT defects observed in HIV infected cells or cells that over-express Rev. Watts et al. hypothesized Rev interacts with MTs by a mechanism shared with Kinesin-13 (Kin13) proteins owing to the presence of a shared amino acid sequence. Kin13 proteins are potent MT depolymerizing agents affecting MT behavior during mitosis. To test this hypothesis, point mutations were introduced into Rev substituting amino acids shared with Kin13. In this study, eight mutant Rev proteins (T34A, …

Light-Limited Access To Fructose Alters Metabolic Function And Adipose Tissue Catecholaminergic Activity In Mice, Naima Salem Rodwan

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Consumption of high levels of fructose produces glucose intolerance, sympathetic nervous system activation, and renal dysfunction. A diet high in fructose may play a role in the worldwide epidemic of obesity and diabetes. The aim of this study was to test the hypothesis that time (light/dark) limited access to fructose influences body fat, metabolic function, and adipose tissue catecholaminergic activity in mice. Male C57BL/6 mice were given standard chow and assigned to one of three groups: Control (n=10, water 24h); FL (n=11, 10% fructose solution during 12h light period); and FD (n=10, 10% fructose solution during 12h dark period). Metabolic …

The Role Of Plant-Soil Feedback In Exotic Plant Invasion: Soil Type, Biotic Or Abiotic Factors?, Kelly Dawn Schradin

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Plants alter soil characteristics in many ways causing changes in their subsequent growth resulting in either positive or negative feedback on their own fitness. Plants in their native ranges typically experience negative feedback from natural enemies, while feedback is often positive in invaded ranges where they escape enemies, experience new beneficial mutualisms, or bring with them a novel biochemical weapon. I conducted a fully factorial greenhouse experiment to examine plant-soil feedback in the invasive shrub Lonicera maackii and whether or not positive feedback may contribute to its successful invasion in southern Ohio. I also investigated whether the sign and strength …

Expression Of Inflamatory Response Genes In Ferrets Challenged With H5n1 Avian Influenza Virus, Brock M. Miniard

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Influenza A H5N1 has emerged as a potential to be the next pandemic influenza. Ferrets are a promising model of H5N1 infection because the disease progression is similar to that known in humans, however reagents to characterize infections in ferrets are few. We developed real-time PCR assays for the ferret cytokines IL-1β, IL-2, IL-4, IL-8, IL-12p40, TNF-α, IFN-α, IFN-β, IFN-γ, TGFB-β1, and TGFB-β2. Forty-eight ferrets were vaccinated with either one of three vaccine formulations (Vaccine 1, Vaccine 2, or Vaccine 3), or saline (0.9% NaCl), then challenged intranasally with a lethal dose of H5N1 strain A/Vietnam/1203/04. Cytokine assays were then …

Examining The Relationship Between Garlic Mustard (Alliaria Petiolata) And European Earthworms, Alexandra M. Zelles

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Our goal was to characterize the interactive feedback between garlic mustard (Alliaria petiolata) and European earthworm species in southwest Ohio. Earthworm community composition, abundance and biomass were compared between 0.1 m² plots of garlic mustard, cut-leaved toothwort (Cardamine concatenate), wild ginger (Asarum canadense), or no plant cover. Exotic earthworms were present in the study site. Earthworm abundance and biomass did not correlate with garlic mustard percent cover. There was a greater density of earthworms in the fall than in the spring. Worm abundance differed between garlic mustard and wild ginger plots and cut-leaved …

Elucidating Transcription Factor Regulation By Tcdd Within The Hs1,2 Enhancer, Sharon D. Ochs

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Immunoglobulin heavy chain (IgH) expression and Ig secretion is inhibited by the environmental contaminant 2,3,7,8-tetrachlorodibenzo-ρ-dioxin (TCDD). Within the IgH gene, the 3' IgH regulatory region (3'IgH RR) has been identified as a transcriptional target of TCDD. The 3'IgH RR, which in part regulates transcription of the IgH gene, is composed of four enhancers in the mouse: hs3a; hs1,2; h3b; hs4 and three enhancers in the human: hs3a; hs1,2; hs4. In humans the hs1,2 enhancer has an invariant sequence (IS) containing a DRE, NF-κB, NF1 and AP-1 binding site. Also, the enhancer has an AP1.ETS and Oct …

Exploration Of Ypel3 Response To Hormones And Ability To Induce Senescence, Joseph E. Rotsinger

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p53 activation through different cellular senescence pathways can trigger cell cycle arrest via regulation of p53 target genes. One such target gene is YPEL3 which is expressed upon binding of tumor suppressor protein p53 at its p53 binding sites (Kelley, 2010). The ability of p53 to induce YPEL3 gene expression led to the discovery that YPEL3 is one of several p53 target genes which induce cellular senescence (Kelley, 2010). Additionally YPEL3 can be regulated independently of p53 by estrogen signaling through estrogen receptor α (Tuttle, 2011). The loss of estrogen receptor α or removal of estrogen induces YPEL3 gene expression …

Establishment Of A Quiescent Infection Of Hsv-1 In L929 Fibroblasts Using A Mitotic Inhibitor And Ifn-Γ, Neelam V. Shinde

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The goal for this study was to determine if a quiescent infection of HSV-1 could be induced in murine fibroblasts L929 by treating them with the anti-mitotic agent 5-fluoro 2'deoxy uridine (FUDR) alone and with interferon-γ. Since neurons are post-mitotic and exhibit a lower metabolic rate than other cells, fibroblasts were treated with FUDR to induce a post-mitotic state. The cell cycle arrest of fibroblasts would decrease the thymidylate metabolism and impair HSV-1 replication. An evaluation of cytopathic effects of FUDR was used to determine the optimal concentration which arrests cell growth and inhibits viral replication. Image J program developed …

Effects Of Conductivity And Fish Grazing On Alkaline Phosphatase Activity Of Littoral Periphyton, Samuel A. Drerup

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Littoral periphyton plays an important role in whole lake primary production and is a substantial contributor to whole lake energy budgets in moderately deep oligotrophic lakes. Under conditions of nutrient deficiency, members of the periphyton community supplement the available nutrient pool through the production of intracellular and extracellular enzymes. These enzymes are used to convert bio-unavailable forms of nitrogen and phosphorous to forms usable by the periphyton community. In this paper, I will investigate how conductivity and fish grazing affect periphyton production and phosphorus acquisition in both field and laboratory experiments.

Evaluation Of Cm-2,525 As A Neuroprotectant Against Sarin: A Comparison With 8-Oh-Dpat, Amanda R. Furman

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Exposure to the chemical warfare agent sarin produces long term neurological deficits. The long term medical consequences could be averted with the development of neuroprotectants to preserve brain function. In our mouse model, the combination of the carboxylesterase inhibitor 2-(o-cresyl)-4H-1:3:2-benzodioxaphosphorin-2-oxide (CBDP) with the organophosphorus (OP) nerve agent sarin was used to render mice more sensitive to poisoning and reduce the amount of hydrogen fluoride cleaved from sarin during binding to esterases. Since carboxylesterase acts as a scavenger, reducing the levels causes sarin to have a greater inhibition effect on acetylcholinesterase. These smaller doses permitted the use of doses similar to …

Q-Ve-Oph, A Control Caspase Inhibitor For Analyzing Neuronal Death, Rebecca L. Bricker

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Sarin is a neurotoxin that has been used in terrorist attacks in Japan and is a potential bioterrorist weapon. It induces seizures by affecting the regulation of neurotransmitters in the brain. Seizures are directly correlated to neuronal damage. Two types of neuronal damage that can occur are apoptosis and necrosis. One of the main regions of the brain where neuronal death occurs is the hippocampus, which is involved in memory. Victims showed chronic decline of memory loss 3 and 7 years after the Tokyo's terrorist attack incident. There are treatments available that can break down sarin or can block the …

Hcn1 Immunoreactivity Of Α-Motoneurons Following Peripheral Nerve Injury, Saif Ahmed

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Peripheral axotomy causes significant alterations in intrinsic motoneuron activity and excitability. Despite successful reinnervation of peripheral targets after injury, the recovery of motor function is incomplete. Following axotomy in the adult cat, there is an increase in the afterhyperpolarization (AHP) of the action potential in specific motoneuron types. AHP duration can be inversely correlated with the amount of Ih or sag current and further shaped through SK (Small conductance calcium-activated potassium) channel currents (Gustafsson and Pinter, 1985; Kuno, Miyata, and Muñoz-Martinez, 1974a). Hyperpolarizing-cyclic nucleotide gated (HCN) channels underlie sag currents and are critical to neuronal function by their unique property …

The Effects Of Enteropathogenic And Commensal Escherichia Coli On Tight Junction Permeability, Hilary Kaye Allen

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The intestinal mucosa maintains a barrier between materials from the external environment and the internal environment of the host. Disruption of the gut wall integrity is involved in the development of various intestinal diseases, such as Irritable Bowel Syndrome (IBS) and Crohn's disease. The intestinal mucosa is lined with epithelial cells that are connected by tight junctions, the intercellular junctions that form a selectively permeable barrier between paracellular pathways. Enteric pathogens, such as enteropathogenic Escherichia coli (EPEC), can disrupt the tight junctions of epithelial cells by altering the cellular cytoskeleton or by directly affecting tight junction proteins. Commensal Escherichia coli …

Comparing Two Different Statins In A Delayed Pharmacological Treatment For Ischemic Stroke, Kailyn M. Hagerty

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Stroke is one of the leading causes of death and the top cause of long term disability. Currently there exists no standardized treatment of care for ischemic stroke patients during the days following stroke other than giving aspirin. A drug combination containing fluoxetine, one of two statins, and ascorbic acid was used to try and improve motor function following strokes in rats. Endothelin induced stroke survival surgery was performed on the right cortex of Long Evans and Sprague Dawley rats. Functional tests were completed pre- and post-surgery and immunohistochemistry was carried out to analyze infarct volume, angiogenesis, and the glial …

Mhc Class I Expression In Murine Fibroblast And Keratinocyte Cell Lines During The First Twenty-Four Hours Of Infection With Herpes Simplex Virus-1 (Hsv-1), Prasanthi Kumchala

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The hypothesis of this study is: HSV-1 infection of murine fibroblasts and keratinocytes inhibits expression of MHC class I molecules during first 24 hours of infection. IFN-γ pretreatment of fibroblasts protected the cells from virus-induced inhibition of MHC class I expression, but did not protect keratinocytes. Herpesviruses are known for their ability to establish persistent infections. Herpesviruses exert many different ways to suppress host defense mechanisms. One such way is by down regulating expression of the major histocompatibility complex I (MHC I) molecules in infected cells. Epidermal cells such as keratinocytes are the major sites for herpes simplex virus type …

The Effects Of Hsv-1 Challenge On Polarized Murine Macrophages: An In Vitro Model Using The J774a.1 Murine Macrophage Cell Line, Adam Craig Reichard

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In our current study we examined the effects of HSV-1 challenge on J774A.1 macrophages polarized to either a proinflammatory (M1) or anti-inflammatory (M2) phenotype. Polarized J774A.1 macrophages were characterized using CD14-CD86 and SOCS1-SOCS3 expression levels. SOCS proteins are a family of proteins that are capable of inhibiting cytokine-signaling pathways. HSV-1 up regulates expression of SOCS1 protein levels in infected cells, inhibiting the ability of infected cells to produce proinflammatory products (Nowoslawski Akhtar and Benveniste, 2011). This study shows that signals within the microenvironment play a greater role in macrophage polarization, and SOCS1-SOCS3 expression levels, than does HSV-1 challenge. M1 macrophages …

Use Of Phage Display Libraries To Select For B-Cell Receptor-Specific Peptides Of Chronic Lymphocytic Leukemia Cells, Richard M. Chou

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Peptides with high affinity to the B-cell receptor (BCR) fused to a toxin could be an effective therapy for Chronic Lymphocytic Leukemia (CLL) patients. We screened captured BCR of a CLL patient with peptides from 7 and 12-mer phage display libraries, using two strategies. Lymphocytes from two patients diagnosed with CLL expressing two different unmutated VH genes (VH 1-3 and VH4-34, respectively) were used. Membrane BCRs were obtained from patient CLL cells by lysis, identified by western blot, semi-quantified and screened with phage libraries. The first strategy involved using patient VH4-34 BCRs which were captured using goat anti-human IgM to …

Powdery Mildew (Erysiphe Cruciferarum) Affects The Allelopathic And Competitive Abilities Of Invasive Garlic Mustard (Alliaria Petiolata), Andrew Russell Officer

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Garlic mustard (Alliaria petiolata) has been previously found to be significantly affected negatively by powdery mildew (Erysiphe cruciferarum). While we could not significantly corroborate those findings we did find evidence that E. cruciferarum does inhibit A. petiolata's allelopathic and competitive effects which benefits some target neighbor species such as Impatiens capensis and Elymus canadensis. We also found that the inhibition of A. petiolata by E. cruciferarum had negative consequences on another neighboring invasive species (Lonicera maackii) compared to those grown next to uninfected A. petiolata. Acer saccharum, a slow-growing species had no effect between neighbors. Sterilization treatments had variable effects …

Trading Resistance For Vigor: A Potential Mechanism For Invasion By The Asian Grass Microstegium Vimineum, Matthew Dickinson

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The Evolution of Increased Competitive Ability (EICA) hypothesis predicts that subsequent to introduction, non-native plants escape enemies and selection favors genotypes that invest more in growth and reproduction and less in resistance. Here, I evaluated if the invasive Asian grass Microstegium vimineum has developed decreased resistance in its introduced range of the eastern US, as predicted by the EICA hypothesis. Asian and US genotypes were evaluated for differences in enemy damage, leaf toughness, specific leaf area (SLA), and flowering in a common garden experiment. Leaf damage, SLA, and flower production were greater and leaf toughness lower in US plants. These …

Regulation Of Mitosis By Nuclear Speckle Proteins, Keshia Nicole Torres-Munoz

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Mitosis is an intricate process that is monitored by multiple cell cycle regulator proteins. Errors in mitotic regulation have been linked to many types of cancer. Recently, evidence suggests an indirect regulation of mitosis by nuclear speckle proteins. Nuclear speckles are one of the multiple compartments found in the mammalian cell nucleus. They serve as assembly compartments for premRNA processing factors. Mass spectrometry analysis of purified nuclear speckles revealed 33 novel proteins including Son and TRAP150. The aim of my study was to determine how Son and TRAP150 can directly or indirectly impact mitosis. We and others recently reported that …

Enhanced Angiotensin Ii-Induced Cardiac And Aortic Remodeling In Ace2 Knockout Mice, Mahmoud Alghamri

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Activation of the renin angiotensin system (RAS) is a well-known driving force, governing the aggravation and progression of CVD and associated target organ damage. Angiotensin (Ang) converting enzyme 2 (ACE2), a new member within the RAS family, was first cloned from the ventricle of patient with heart failure. ACE2 was shown to be upregulated under pathological conditions. We hypothesize that loss of ACE2 negatively affects cardiac function and exacerbates hypertrophic cardiomyopathy and aortic remodeling in response to Ang II. Eight weeks old ACE2 knock out (KO) and wild type (WT) mice were infused with Ang II s.c. (1000 ng/kg/min for …

The Use Of Forelimb Asymmetry Functional Tests To Determine Motor Recovery With Various Drug Treatments Following Endothelin-Induced Stroke, Kelly Rebecca Leach

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Our laboratory has found increased functional recovery post-stroke with Long Evans rats that received a combination of fluoxetine, simvastatin, and vitamin C. The objectives of this study were to test that treatment and other drug combinations on Sprague Dawley rats, to determine whether Forelimb Asymmetry is a reliable behavioral measure for stroke studies, and to determine the most optimal surgical procedure on Sprague Dawley rats. There were more significant increases in functional recovery observed on the Forelimb Asymmetry than on Montoya Staircase behavioral tests, but there were no significant differences between treatment groups and control. The largest number of rats …

Study Of Microrna-34a Mediated Post Transcriptional Regulation Of Mdm4, Pooja P. Mandke

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MDM4 is an important negative regulator of the tumor suppressor p53. In normal unstressed cells the activity of p53 is kept under control by MDM4 and its homologue MDM2. MDM4 is said to possess oncogenic potential based on the evidence of its overexpression in many cancers. Until recently it was believed MDM4 is constitutively transcribed; however a decrease in full length MDM4 in response to genotoxic stress was observed paving way for exploring the mechanism responsible for this.

It was observed miR-34a a member of the miR34 family which is a direct transcriptional targets of p53 could have a potential …

Role Of Na+K+2cl- Co-Transporters In Insulin Secretion, Saeed Alshahrani

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The objective of this study is to investigate the role of intracellular chloride concentration ([Cl^-]i) in insulin secretion in vivo and in vitro.

The maintenance of high [Cl^-]i in insulin-secreting pancreatic β-cells plays an important role in glucose-induced depolarization and insulin secretion. Insulinotropic glucose concentrations i.e., >6 mM induces electrical activity in β-cells which results in insulin secretion. This electrical activity is mainly due to electrogenic efflux of Cl^- from β-cells through volume-regulated anion channels (VRAC). The solute carrier family 12 group A member 1 (Slc12a1) and 2 (Slc12a2) encode several splice variants of the …

Suppressor Of Cytokine Signaling (Socs) 1 & 3 Expression In Hsv-1- Infected And Interferon-Γ-Treated Neuro-2a Cells, Melinda Jones

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This study examined the effects of HSV-1 infection and IFN-γ treatment on Neuro-2A cells. HSV-1 induces expression of SOCS1 and SOCS3 in infected cells, inhibiting the ability of these cells to produce the pro inflammatory, antiviral cytokine IFN-γ (Nowoslawski and Benveniste, 2011). SOCS1 and SOCS3 levels were determined in IFN-γ-treated cells, virus-infected cells, and cells that were both IFN-γ-treated and virus-infected. Results were compared with untreated, uninfected control cells. Flow cytometry data analysis showed a slight decrease in SOCS1 and SOCS3 protein levels in cells treated with IFN-γ for 6 hours compared to control cells. A significant decrease in SOCS1 …

The Construction Of A Plasmid For Detecting The Pathway Of Arginine Metabolism In Human Macrophages: A Real-Time Assessment Of Macrophage Polarity, Benjamin A. Holmes

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Macrophages are "first responders", innate immune system cells which quickly arrive to a site of infection and injury, consuming cell debris and foreign matter and recruiting other immune system cells to the area. While historically they have been thought to react uniformly to all challenges, the discovery of toll-like receptors has shown that macrophages actually work closely with the adaptive immune system in fine-tuning the immune response. Furthermore, it has recently been discovered that macrophages can become polarized to one of two subtypes-M1 or M2.

M1 macrophages are efficient producers of reactive oxygen species, nitrogen intermediates, and inflammatory cytokines. They …

Lipase-Kinase Associations Involving Pld2, Jak3 And Fes That Underlie Cancer Cell Proliferation And Invasion, Qing Ye

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Phospholipase D (PLD) is an enzyme that breaks down phospholipids in the cell membrane. It has been suggested that PLD may play a role during cell proliferation and cell invasion of cancer cells. The objective of this thesis was to define new molecular signaling pathways in which PLD2 might be involved in terms of cell proliferation (first part) and cell invasion (second part). To this, I compared molecular and biochemical aspects between untransformed cell lines with highly invasive, transformed breast cancer cells. In the first part, I investigated the interaction of two tyrosine kinases with PLD2 and the effect of …

Expressing Human Orai3 In Insect Cells For Pharmacological Studies, Orville R. Bennett

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The Orai3 protein forms a Ca2+ channel with a pharmacological profile distinct from its close relative, the store-operated Ca2+ channel Orai1. Though closely related to Orai1, the function of Orai3 in humans is still unclear. This study attempts to contribute to the body of knowledge by undertaking a pharmacological analysis of Orai3 in insect cells. We describe here the creation of a vector capable of expressing the mammalian Orai3 gene on an insect background. We demonstrate the ability to induce gene expression of Orai3 in these insect cells, and then assess the effectiveness of this system by characterizing the pharmacological …