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Expression Of Rna Nanoparticles Based On Bacteriophage Phi29 Prna In Escherichia Coli And Bacillus Subtilis, Le Zhang

Theses and Dissertations--Pharmacy

Currently, most of the RNAs used in lab research are prepared by in vitro transcription or chemical synthesis, which can be costly. In vivo expression in bacterial cells is another approach to RNA preparation that allows large scale production at a lower cost. However, there are some obstacles in bacterial expression, including RNA degradation in host cell, as well as RNA extraction and purification. tRNA and 5S RNA have been reported as scaffolds to circumvent the degradation problem. These scaffolds can not only make the RNA product survive in the cell but also increase the stability after extraction.

The packaging …

Early Stage Drug Treatment That Normalizes Proinflammatory Cytokine Production Attenuates Synaptic Dysfunction In A Mouse Model That Exhibits Age-Dependent Progression Of Alzheimer's Disease-Related Pathology, Adam D. Bachstetter, Christopher M. Norris, Pradoldej Sompol, Donna M. Wilcock, Danielle Goulding, Janna H. Neltner, Daret St. Clair, D. Martin Watterson, Linda J. Van Eldik

Sanders-Brown Center on Aging Faculty Publications

Overproduction of proinflammatory cytokines in the CNS has been implicated as a key contributor to pathophysiology progression in Alzheimer's disease (AD), and extensive studies with animal models have shown that selective suppression of excessive glial proinflammatory cytokines can improve neurologic outcomes. The prior art, therefore, raises the logical postulation that intervention with drugs targeting dysregulated glial proinflammatory cytokine production might be effective disease-modifying therapeutics if used in the appropriate biological time window. To test the hypothesis that early stage intervention with such drugs might be therapeutically beneficial, we examined the impact of intervention with MW01-2-151SRM (MW-151), an experimental therapeutic that …

Session 1c, Kentucky Water Resources Research Institute, University Of Kentucky

Kentucky Water Resources Annual Symposium

• The Effects of Prescribed Fire on Amphibian and Reptile Diversity in an Oak-Grassland Restoration Area, Robert Knopp and Howard Whiteman, Dept Biological Sciences and Watershed Studies, Murray State University
• Results of a Data Evaluation to Establish Priority Remediation Areas for Dry Weather Fecal Contamination in a Karst Influenced Watershed, Steve Evans and others, Third Rock Consultants, Lexington, KY
• Development of In-House Methods for High-Throughput DNA Extraction, Kimberly L. Cook and LorraBelle Hill, USDA-ARS, Bowling Green, KY
• Validation of the Sanitary Category Value Model (SCV) for the Identification of Leaking Sewer Lines: A Study of the West Hickman …

Proceedings Of 2012 Kentucky Water Resources Annual Symposium, Kentucky Water Resources Research Institute, University Of Kentucky

Kentucky Water Resources Annual Symposium

This symposium was planned and conducted as a part of the state water resources research institute annual program that is supported by Grant/Cooperative Agreement Number G11AP20081 from the United States Geological Survey. The contents of this proceedings document and the views and conclusions presented at the symposium are solely the responsibility of the individual authors and presenters and do not necessarily represent the official views of the USGS or of the symposium organizers and sponsors. This publication is produced with the understanding that the United States Government is authorized to reproduce and distribute reprints for government purposes.

Examining Chronic Non-Cancer Pain Among A Sample Of Individuals In Opioid Treatment Programs, Erin Stevenson

Theses and Dissertations--Social Work

National rates of chronic non-cancer pain (CNCP) are rising alongside increasing reports of prescription opioid abuse and mortality. Associations between the rise in CNCP and in opioid abuse seem logical, yet research on CNCP among individuals with opioid dependence is currently limited due to the complicated nature of comorbid conditions in research and treatment. This study aims to expand the CNCP knowledge base by responding to the question: Do individuals with CNCP participating in an opiate treatment program have better or worse treatment outcomes than individuals without CNCP?

This study used a secondary dataset including 483 adults from Kentucky’s Opiate …

Effect Of Variable Transmission Rate On The Dynamics Of Hiv In Sub-Saharan Africa, Diego F. Cuadros, Phillip H. Crowley, Ben Augustine, Sarah L. Stewart, Gisela García-Ramos

Biology Faculty Publications

BACKGROUND: The cause of the high HIV prevalence in sub-Saharan Africa is incompletely understood, with heterosexual penile-vaginal transmission proposed as the main mechanism. Heterosexual HIV transmission has been estimated to have a very low probability; but effects of cofactors that vary in space and time may substantially alter this pattern.

METHODS: To test the effect of individual variation in the HIV infectiousness generated by co-infection, we developed and analyzed a mathematical sexual network model that simulates the behavioral components of a population from Malawi, as well as the dynamics of HIV and the co-infection effect caused by other infectious diseases, …

Viability Of A Controllable Chaotic Micromixer Through The Use Of Titanium-Nickel Shape Memory Alloy, David Ryan Lilly

Theses and Dissertations--Mechanical and Aerospace Engineering

Microfluidic devices have found applications in a number of areas, such as medical analysis, chemical synthesis, biological study, and drug delivery. Because of the small channel dimensions used in these systems, most microchannels exhibit laminar flow due to their low Reynold’s number, making mixing of fluids very challenging. Mixing at this size scale is diffusion-limited, so inducing chaotic flow patterns can increase the interface surface area between two fluids, thereby decreasing overall mixing time.

One method to create a chaotic flow within the channel is through the introduction of internal protrusions into the channel. In such an application protrusions that …

The Role Of Co-Infection In The Spread Of Hiv In Sub-Saharan Africa, Diego Fernando Cuadros

Theses and Dissertations--Biology

The cause of the high HIV prevalence in sub-Saharan Africa is incompletely understood, with heterosexual penile-vaginal transmission proposed as the main mechanism. Heterosexual HIV transmission has a very low probability; further, a single estimation of heterosexual probability of HIV transmission fails to reproduce the variation associated with important biological cofactors. In particular, studies of HIV incidence suggest that co-infection with other infectious diseases influence the HIV transmission, and therefore might substantially vary the pattern of the spread of the infection. To assess the effect of co-infection on the spread of HIV, I developed and analyzed several mathematical and statistical models …

Time's Arrow Flies Like A Bird: Two Paradoxes For Avian Circadian Biology, Vincent M. Cassone, Jiffin K. Paulose, Melissa G. Whitfield-Rucker, Jennifer L. Peters

Biology Faculty Publications

Biological timekeeping in birds is a fundamental feature of avian physiology, behavior and ecology. The physiological basis for avian circadian rhythmicity has pointed to a multi-oscillator system of mutually coupled pacemakers in the pineal gland, eyes and hypothalamic suprachiasmatic nuclei (SCN). In passerines, the role of the pineal gland and its hormone melatonin is particularly important. More recent molecular biological studies have pointed to a highly conserved mechanism involving rhythmic transcription and translation of "clock genes". However, studies attempting to reconcile the physiological role of pineal melatonin with molecular studies have largely failed. Recent work in our laboratory has suggested …

Genic Regions Of A Large Salamander Genome Contain Long Introns And Novel Genes, Jeramiah J. Smith, Srikrishna Putta, Wei Zhu, Gerald M. Pao, Inder M. Verma, Tony Hunter, Susan V. Bryant, David M. Gardiner, Timothy T. Harkins, S. Randal Voss

Biology Faculty Publications

BACKGROUND: The basis of genome size variation remains an outstanding question because DNA sequence data are lacking for organisms with large genomes. Sixteen BAC clones from the Mexican axolotl (Ambystoma mexicanum: c-value = 32 x 10(9) bp) were isolated and sequenced to characterize the structure of genic regions.

RESULTS: Annotation of genes within BACs showed that axolotl introns are on average 10x longer than orthologous vertebrate introns and they are predicted to contain more functional elements, including miRNAs and snoRNAs. Loci were discovered within BACs for two novel EST transcripts that are differentially expressed during spinal cord regeneration and skin …

Differential Gene Expression Associated With Postnatal Equine Articular Cartilage Maturation, Michael J. Mienaltowski, Liping Huang, Arnold J. Stromberg, James N. Macleod

Veterinary Science Faculty Publications

BACKGROUND: Articular cartilage undergoes an important maturation process from neonate to adult that is reflected by alterations in matrix protein organization and increased heterogeneity of chondrocyte morphology. In the horse, these changes are influenced by exercise during the first five months of postnatal life. Transcriptional profiling was used to evaluate changes in articular chondrocyte gene expression during postnatal growth and development.

METHODS: Total RNA was isolated from the articular cartilage of neonatal (0-10 days) and adult (4-5 years) horses, subjected to one round of linear RNA amplification, and then applied to a 9,367-element equine-specific cDNA microarray. Comparisons were made with …

A Novel Class Of Immunoproteasome Catalytic Subunit Lmp2 Inhibitor And Its Therapeutic Potentials In Cancer, Yik Khuan (Abby) Ho

University of Kentucky Doctoral Dissertations

The immunoproteasome, known to play an important role in MHC class I antigen processing and presentation, have been linked to neurodegenerative diseases and hematological cancers. However, the pathophysiological functions of the immunoproteasome in these diseases are still not very well established. This can be attributed mainly to the lack of appropriate molecular probes that selectively target the immunoproteasome catalytic subunits. Herein, we report the development of a small molecular inhibitor (AM) that selectively targets the major catalytic subunit, LMP2, of the immunoproteasome. We show that the compound covalently modifies the LMP2 subunit with high specificity in human prostate …

The Interaction Of Haptic Imagery With Haptic Perception For Sighted And Visually Impaired Consumers, Shannon Bridgmon Rinaldo

University of Kentucky Doctoral Dissertations

Consumers evaluate products in the market place using their senses and often form mental representations of product properties. These mental representations have been studied extensively. Imagery has been shown to interact with perception within many perceptual modalities including vision, auditory, olfactory, and motor. This dissertation draws on the vast visual imagery literature to examine imagery in the haptic, or touch, modality. Two studies were undertaken to examine the relationship between haptic imagery and haptic perception The first study is based on studies from cognitive psychology that have used similar methods for examining visual imagery and visual perception. In study 1, …

Methods Development In Qualitative And Quantitative Proteomics, Ting Liu

University of Kentucky Doctoral Dissertations

Proteomics based on liquid chromatography coupled to mass spectrometry has developed rapidly in the last decade and become a powerful tool for protein mixtures analysis. LC-MS based proteomics involves four steps, sample preparation, liquid chromatography, mass spectrometry and bioinformatics. Improvements in each step have extended its applications to new biological research areas. This dissertation mainly focuses on method developments in both qualitative and quantitative proteomics.

The first part of this dissertation focuses on qualitative analysis of T. gondii Parasitophorous Vacuole Membrane (PVM) proteins, which is very important for T. gondii’s survival. The hypothesis of this study is that proteomic …

A Non-Circadian Role For Clock-Genes In Sleep Homeostasis: A Strain Comparison, Paul Franken, Ryan Thomason, H. Craig Heller, Bruce F. O'Hara

Biology Faculty Publications

BACKGROUND: We have previously reported that the expression of circadian clock-genes increases in the cerebral cortex after sleep deprivation (SD) and that the sleep rebound following SD is attenuated in mice deficient for one or more clock-genes. We hypothesized that besides generating circadian rhythms, clock-genes also play a role in the homeostatic regulation of sleep. Here we follow the time course of the forebrain changes in the expression of the clock-genes period (per)-1, per2, and of the clock-controlled gene albumin D-binding protein (dbp) during a 6 h SD and subsequent recovery sleep in three inbred strains of mice for which …

A Simple Array Platform For Microrna Analysis And Its Application In Mouse Tissues, Xiaoqing Tang, Jozsef Gal, Xun Zhuang, Wang-Xia Wang, Haining Zhu, Guiliang Tang

Plant and Soil Sciences Faculty Publications

MicroRNAs (miRNAs) are a novel class of small noncoding RNAs that regulate gene expression at the post-transcriptional level and play a critical role in many important biological processes. Most miRNAs are conserved between humans and mice, which makes it possible to analyze their expressions with a set of selected array probes. Here, we report a simple array platform that can detect 553 nonredundant miRNAs encompassing the entire set of miRNAs for humans and mice. The platform features carefully selected and designed probes with optimized hybridization parameters. Potential cross-reaction between mature miRNAs and their precursors was investigated. The array platform was …

Energizing Mirna Research: A Review Of The Role Of Mirnas In Lipid Metabolism, With A Prediction That Mir-103/107 Regulates Human Metabolic Pathways, Bernard R. Wilfred, Wang-Xia Wang, Peter T. Nelson

Sanders-Brown Center on Aging Faculty Publications

MicroRNAs (miRNAs) are powerful regulators of gene expression. Although first discovered in worm larvae, miRNAs play fundamental biological roles-including in humans-well beyond development. MiRNAs participate in the regulation of metabolism (including lipid metabolism) for all animal species studied. A review of the fascinating and fast-growing literature on miRNA regulation of metabolism can be parsed into three main categories: (1) adipocyte biochemistry and cell fate determination; (2) regulation of metabolic biochemistry in invertebrates; and (3) regulation of metabolic biochemistry in mammals. Most research into the 'function' of a given miRNA in metabolic pathways has concentrated on a given miRNA acting upon …

Aging And The Dynorphinergic System: Evaluation Of Memory And Motor Systems In Prodynorphin Knockout Mice, Xuan V. Nguyen

University of Kentucky Doctoral Dissertations

Dynorphins, endogenous peptide neurotransmitters expressed in the central nervous system, have been implicated in diverse pathophysiological processes, including excitotoxicity, chronic inflammation, traumatic injury, cognitive impairment, and motor dysfunction, with significant changes with aging or age-related disease processes. This has led to the hypothesis that the suppression of dynorphin expression would produce beneficial effects on learning and memory and motor function. To assess the phenotypic manifestations of chronic suppression of endogenous dynorphin, knockout (KO) mice lacking the coding exons of the gene encoding the prodynorphin (Pdyn) precursor protein, were tested in a series of behavioral, biochemical, and molecular biological studies. Moderately …

Spontaneous Intracranial Arterial Dissection In The Young: Diagnosis By Ct Angiography, William C. Robertson, Curtis A. Given

Neurology Faculty Publications

BACKGROUND: Spontaneous carotid artery dissections have been rarely reported in children. Diagnosis has traditionally been confirmed by catheter arteriography. More recently diagnosis has been made by magnetic resonance imaging and magnetic resonance angiography; however the sensitivity of these techniques has yet to be determined. The authors are unaware of reports of carotid dissection confirmed by dynamic computed tomography (computerized tomographic arteriography) in the young.

CASE PRESENTATION: We recently evaluated a fourteen year-old male following the development of transient neurologic symptoms. There was no antecedent illness or trauma. Dynamic computed tomography revealed an intracranial dissection involving the supraclinoid segment of the …

Synthases, Joseph Chappell, Kathleen R. Manna, Joseph P. Noel, Courtney M. Starks

Pharmaceutical Sciences Faculty Patents

Novel synthases and the corresponding nucleic acids encoding such synthases are disclosed herein. Such synthases possess an active site pocket that includes key amino acid residues that are modified to generate desired terpenoid reaction intermediates and products. Synthase modifications are designed based on, e.g., the three-dimensional coordinates of tobacco 5-epi-aristolochene synthase. with or without a substrate bound in the active site.

The Role Of Tombusvirus Replicase Proteins And Rna In Replicase Assembly, Replication And Recombination, Zivile Sliesaraviciute Panaviene

University of Kentucky Doctoral Dissertations

Tombusviruses are single, positive strand RNA viruses of plants, often associated with parasitic defective interfering (DI) RNAs. Two viral- coded gene products, namely p33 and p92, are required for tombusvirus replication. The overlapping domains of p33 and p92 contain an arginine/proline-rich (RPR) RNA binding motif. In this study, the role of RPR motif and viral RNA in tombusvirus replication and recombination, as well as involvement of viral RNA in tombusvirus replicase assembly was examined. Using site-directed mutagenesis I generated a series of RPR mutants of Cucumber necrosis tombusvirus (CNV). Analysis of RPR mutants defined that wild type RPR motif, especially …

Synthases, Joseph Chappell, Kathleen R. Manna, Joseph P. Noel, Courtney M. Starks

Pharmaceutical Sciences Faculty Patents

Novel synthases and the corresponding nucleic acids encoding such synthases are disclosed herein. Such synthases possess an active site pocket that includes key amino acid residues that are modified to generate desired terpenoid reaction intermediates and products. Synthase modifications are designed based on, e.g., the three-dimensional coordinates of tobacco 5-epi-aristolochene synthase, with or without a substrate bound in the active site.

Synthases, Joe Chappell, Kathleen R. Manna, Joseph P. Noel, Courtney M. Starks

Pharmaceutical Sciences Faculty Patents

Novel synthases and the corresponding nucleic acids encoding such synthases are disclosed herein. Such synthases possess an active site pocket that includes key amino acid residues that are modified to generate desired terpenoid reaction intermediates and products. Synthase modifications are designed based on, e.g., the three-dimensional coordinates of tobacco 5-epi-aristolochene synthase, with or without a substrate bound in the active site.

Synthases, Joe Chappell, Kathleen R. Manna, Joseph P. Noel, Courtney M. Starks

Pharmaceutical Sciences Faculty Patents

Novel synthases and the corresponding nucleic acids encoding such synthases are disclosed herein. Such synthases possess an active site pocket that includes key amino acid residues that are modified to generate desired terpenoid reaction intermediates and products. Synthase modifications are designed based on, e.g., the three-dimensional coordinates of tobacco 5-epi-aristolochene synthase, with or without a substrate bound in the active site.

Synthases, Joseph Chappell, Kathleen R. Manna, Joseph P. Noel, Courtney M. Starks

Pharmaceutical Sciences Faculty Patents

Novel synthases and the corresponding nucleic acids encoding such synthases are disclosed herein. Such synthases possess an active site pocket that includes key amino acid residues that are modified to generate desired terpenoid reaction intermediates and products. Synthase modifications are designed based on, e.g., the three-dimensional coordinates of tobacco 5-epi-aristolochene synthase with or without a substrate bound in the active site.

Methods Of Making Modified Polypeptides, Joseph Chappell, Kathleen R. Manna, Joseph P. Noel, Courtney M. Starks

Pharmaceutical Sciences Faculty Patents

Novel polypeptides and the corresponding nucleic acids encoding such polypeptides are disclosed herein. The invention provides methods of making modified polypeptides by altering one or more amino acid residues involved in the active site of a preselected polypeptide.

Water Quality In The Kentucky River Basin, Daniel I. Carey

Information Circular--KGS

Data gathered up to 1990 suggest that water pollution problems existed throughout the Kentucky River Basin. Fecal coliform bacteria in streams was a widespread problem because of the inadequate treatment of municipal wastes, failing septic systems, and agricultural runoff. Iron, lead, manganese, mercury, and silver exceeded State standards and Federal guidelines for drinking water and aquatic life at most of the sample sites for a majority of samples. Aquatic life in many smaller streams in the Knobs region was reduced by chloride discharges from oil and gas operations, according to the Kentucky Division of Water. Organic enrichment and high nutrient …

A Chronicle Of The Kentucky Transportation Research Program, Robert C. Deen, Ronald D. Hughes

Kentucky Transportation Center Research Report

The basis for the Kentucky Transportation Research Program (KTRP) was established more than 45 years ago and some of the current staff members have over 35 years experience in transportation research. Over the years, principal investigators have developed expertise in major areas of transportation and have made significant contributions in highway safety, pavement design and performance, embankment analyses, fatigue detection in bridges, noise abatement, traffic control and operations, voidless concrete, pavement texture and skid resistance, structural design and analyses of culverts, traffic forecasting, and numerous other areas.

The Transportation Research Building houses several fully equipped laboratories. Special effort was made …

Kentucky's Ipm Program For Alfalfa, Chris M. Christensen

Kentucky Alfalfa and Stored Forage Conference

Integrated Pest Management (IPM) is a concept that encourages the use of a variety of strategies in the management of crop pests. It is very important to have an indepth understanding of the crop production system when implementing an IPM program. An IPM program is dynamic requiring a continual evaluation of the status of the crop, the associated pest complex, the effects of the pests and the biological deterrents to those pests. If a pest population gets "out of hand" and begins to cause economic damage, pest management techniques may need to be applied. These pest management measures may be …

Tritium (3h) Radiolabeling Of Protein A And Antibody To High Specific Activity: Application To Cell Surface Antigen Radioimmunoassays, Ronald L. Wilder, Catherine C. Yuen, Subbarao Bondada, Virgil L. Woods, Cornelius B. Alexander, Rose G. Mage

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Staphylococcal protein A and several different immunoglobulins have been radio-labeled to high specific activities (> 106 cpm/pg) by reductive methylation with tritiated (3H) sodium borohydride. The proteins retain excellent functional and antigenic properties. The utility of these reagents in a variety of assays for cell surface antigens is illustrated. The results indicate that this radiolabeling procedure may become the method of choice for many cell surface and solution immunoassays.