Medicine and Health Sciences Commons^™

Studies On The Role Of Acid Sphingomyelinase And Ceramide In The Regulation Of Tace Activity And Tnfα Secretion By Macrophages, Krasimira Rozenova

University of Kentucky Doctoral Dissertations

Acid Sphingomyelinase (ASMase) activity has been proposed to mediate LPS signaling in various cell types. This study shows that in macrophages, ASMase is a negative regulator of LPS-induced TNFα secretion. ASMasedeficient (asm^-/-) mice and isolated peritoneal macrophages produce several fold more TNFα than their wild-type (asm^+/+) counterparts when stimulated with LPS. The mechanism for these differences however is not transcriptional but post-translational.

The TNFα converting enzyme (TACE) catalyzes the maturation of the 26kD precursor (proTNFα) to the active 17kD form (sTNFα). In mouse peritoneal macrophages, the activity of TACE rather than the rate of TNFα mRNA …

Computational Design Of 3-Phosphoinositide Dependent Kinase-1 Inhibitors As Potential Anti-Cancer Agents, Mohamed Diwan Mohideen Abdulhameed

University of Kentucky Doctoral Dissertations

Computational drug design methods have great potential in drug discovery particularly in lead identification and lead optimization. 3-Phosphoinositide dependent kinase-1 (PDK1) is a protein kinase and a well validated anti-cancer target. Inhibitors of PDK1 have the potential to be developed as anti-cancer drugs. In this work, we have applied various novel computational drug design strategies to design and identify new PDK1 inhibitors with potential anti-cancer activity. We have pursued novel structure-based drug design strategies and identified a new binding mode for celecoxib and its derivatives binding with PDK1. This new binding mode provides a valuable basis for rational design of …

Synthesis And Evaluation Of Prouteolysis Taurgeting Chimeras (Protacs): A Potential Chemical Genetic Approach To Breast Cancer Therapy, Kedra C. Cyrus

University of Kentucky Doctoral Dissertations

The use of small molecules to probe the function of proteins is referred to as chemical genetics. The Proteolysis Targeting Chimera (PROTAC) is a chemical genetic tool that contains the ligand for a target protein of interest and the recognition motif for an E3 ubiquitin ligase attached by a linker. The PROTAC is capable of binding to and recruiting specific target proteins to the intracellular degradation system, the ubiquitin proteasome system (UPS). While the approach has had success it has not been optimized to be used on a broader scale.

Optimization efforts focused on elucidating the ideal linker length between …

Role Of Aryl Hydrocarbon Receptor In Chronic Inflammatory Diseases, Violeta Arsenescu

University of Kentucky Doctoral Dissertations

Aryl Hydrocarbon Receptor (AhR) is a ligand-actviated receptor known as the dioxin receptor. Environmental pollutants called dioxin-like toxicants are found in food, cigarette smoke, automobile exhaust and air. Therefore, they could chronically amplify the pathology of numerous chronic inflammatory diseases. AhR is a well known target of these environmental chemicals that disrupt endocrine signaling. By the year 2020, the number of people older than 60 years is expected to top 1 billion. The burden of treating chronic disease is significant both in dollars spent and in lost productivity. The need to identify risk factors for chronic diseases must be evaluated …

The Pharmacogenomics Of Egfr-Dependent Nsclc: Predicting And Enhancing Response To Targeted Egfr Therapy, Justin M. Balko

University of Kentucky Doctoral Dissertations

The introduction of tyrosine kinase inhibitors (TKI) targeting the epidermal growth factor receptor (EGFR) inhibitors to the clinic has resulted in an improvement in the treatment of non small cell lung cancer (NSCLC). However, many patients treated with EGFR TKIs do not respond to therapy. The burden of failed treatment is largely placed on the healthcare field, limiting the effectiveness of EGFR TKIs. Furthermore, responses are hindered by the emergence of resistance. Thus, two questions must be addressed to achieve maximum benefit of EGFR inhibitors: How can patients who will benefit from EGFR TKIs be selected a priori? How …

The Interplay Between The Expression And Functions Of Wnt13 Isoforms During Apoptosis In Bovine Aortic Endothelial Cells, Tao Tang

University of Kentucky Doctoral Dissertations

Wnt proteins are crucial for development/homeostasis by controlling cell fate including apoptosis (Moon RT et al. 1997). Three humanWnt13 isoforms were identified: the secreted Wnt13A, mitochondrial Wnt13B, and nuclear Wnt13C forms; and nuclear Wnt13 had an increased sensitivity to LPS/TNF-induced apoptosis in primary endothelial cells (EC); both Wnt13B and C mRNA contain two start codons (AUG+1 and +74), but the same protein encoded from AUG+74 by Wnt13C was expressed lower than Wnt13B (Struewing IT et al.2006). We hypothesize that during EC apoptosis, the nuclear Wnt13C expression is regulated translationally; nuclear Wnt13 favors apoptosis through regulating the activity/expression of apoptosis-related factors; …

Post Discharge Nausea And Vomiting In Ambulatory Surgical Patients: Incidence And Management Strategies, Jan Odom Forren

University of Kentucky Doctoral Dissertations

Approximately 65% of all surgeries are conducted in the outpatient surgery setting involving more than 35 million patients. Thirty-five to fifty percent of these outpatients will experience post discharge nausea and vomiting (PDNV), nausea and vomiting that occurs after discharge from the health care facility after surgery. A dearth of literature details the problems associated with nausea and vomiting experienced by patients after discharge home from outpatient surgery.

The purposes of this dissertation were to (1) review the current knowledge in the area of post discharge nausea and vomiting; (2) present results of an integrative review of the research literature …

Low Density Lipoprotein Receptor And Alzheimers Disease, Rangaraj K. Gopalraj

University of Kentucky Doctoral Dissertations

Since apoE allele status is the predominant Alzheimers disease (AD) genetic risk factor, functional single nucleotide polymorphisms (SNPs) in brain apoE receptors represent excellent candidates for association with AD. Therefore, three low density lipoprotein receptor (LDLR) SNPs were evaluated by TaqMan allelic discrimination assays for association with AD and I found that certain haplotypes alter the odds of AD. A SNP within LDLR exon 12, rs688, was identified in silico as neutralizing a putative exon splicing enhancer (ESE). Since LDLR is a major apoE receptor in the brain, I hypothesized that rs688 modulates LDLR splicing in neural tissues and associates …

Gender Specific Differences In The Pros And Cons Of Smoking Among Current Smokers In Eastern Kentucky: Implications For Future Smoking Cessation Interventions, Dana A. Hazen, David M. Mannino, Richard Clayton

Preventive Medicine and Environmental Health Faculty Publications

This study investigated gender differences in the perceived “pros” and “cons” of smoking using the constructs of decisional balance (DB) and stage of change from the Transtheoretical Model. The population distribution for stage of change among a population-based, cross-sectional survey of 155 current smokers over 40 years was: precontemplation (22.6%), contemplation (41.9%), preparation (35.5%). Results of stepwise regression models indicated significant gender differences in DB were in the preparation stage of change; scores on the DB measure increased 3.94 points (95% CI: 1.94, 5.93) for male smokers. Interventions targeting the “pros” and “cons” of smoking may need to be gender …

Borderline Personality Disorder: An Overview Of History, Diagnosis And Treatment In Adolescents, Linah Al-Alem, Hatim A. Omar

Pharmacology and Nutritional Sciences Faculty Publications

Borderline personality disorder (BPD) is a cluster B personality disorder. It is characterized by erratic behaviors, emotional instability and one of its hallmarks is self injurious behavior, which starts in adolescence. Patients with BPD are difficult to treat, most have a history of child sexual abuse, about a quarter present with sexual abuse from a caretaker. Although personality disorders are diagnosed only in adults, BPD manifests itself in adolescence in the form of uncontrollable anger, self mutilations, dissociation and other such behaviors. Hence, there is a growing number of scientists discussing the possibility of diagnosing BPD in adolescents. Here, we …

Aluminum Bioavailability From Tea Infusion, Robert A. Yokel, Rebecca L. Florence

Pharmaceutical Sciences Faculty Publications

The objective was to estimate oral Al bioavailability from tea infusion in the rat, using the tracer ²⁶Al. ²⁶Al citrate was injected into tea leaves. An infusion was prepared from the dried leaves and given intra-gastrically to rats which received concurrent intravenous ²⁷Al infusion. Oral Al bioavailability (F) was calculated from the area under the ²⁶Al, compared to ²⁷Al, serum concentration × time curves. Bioavailability from tea averaged 0.37%; not significantly different from water (F = 0.3%), or basic sodium aluminum phosphate (SALP) in cheese (F = 0.1 to 0.3%), but greater than acidic SALP …

Manufactured Aluminum Oxide Nanoparticles Decrease Expression Of Tight Junction Proteins In Brain Vasculature, Lei Chen, Robert A. Yokel, Bernhard Hennig, Michal Toborek

Pharmaceutical Sciences Faculty Publications

Manufactured nanoparticles of aluminum oxide (nano-alumina) have been widely used in the environment; however, their potential toxicity provides a growing concern for human health. The present study focuses on the hypothesis that nano-alumina can affect the blood-brain barrier and induce endothelial toxicity. In the first series of experiments, human brain microvascular endothelial cells (HBMEC) were exposed to alumina and control nanoparticles in dose- and time-responsive manners. Treatment with nano-alumina markedly reduced HBMEC viability, altered mitochondrial potential, increased cellular oxidation, and decreased tight junction protein expression as compared to control nanoparticles. Alterations of tight junction protein levels were prevented by cellular …

Young People's Blood Alcohol Concentration And The Alcohol Consumption City Law, Brazil, Raquel De Boni, Carl G. Leukefeld, Flavio Pechansky

Center on Drug and Alcohol Research Faculty Publications

The paper assesses blood alcohol concentration and risk behaviors for traffic accidents before and after the implementation of a law which prohibits the use of alcoholic beverages on city gas stations. In Porto Alegre, Southern Brazil, young people go out at night and drive to gas station convenience stores to buy alcoholic beverages which are consumed on the premises of parking lots in gas stations. Data were obtained from self-administered questionnaires and breath analyzers in two cross-sectional collections with purposive samples of youngsters in May and July 2006 (n=62, and n=50, respectively). There were no significant differences between the groups …

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 …

Crystallization And Structure Of A Plant Peptide Deformylase, Robert L. Houtz, David W. Rodgers, Lynnette M. A. Dirk, Mark A. Williams

Molecular and Cellular Biochemistry Faculty Patents

This invention relates to the crystal structure of a plant peptide deformylase polypeptide and methods of using the structure to design compounds that modulate the activity of the polypeptide.

Childhood Lymphohematopoietic Cancer Incidence And Hazardous Air Pollutants In Southeast Texas, 1995–2004, Kristina W. Whitworth, Elaine Symanski, Ann L. Coker

CRVAW Faculty Journal Articles

BACKGROUND: Cancer is the second leading cause of death among U.S. children with few known risk factors. There is increasing interest in the role of air pollutants, including benzene and 1,3-butadiene, in the etiology of childhood cancers.

OBJECTIVE: Our goal was to assess whether census tracts with the highest benzene or 1,3-butadiene ambient air levels have increased childhood lymphohematopoietic cancer incidence.

METHODS: Our ecologic analysis included 977 cases of childhood lymphohematopoietic cancer diagnosed from 1995–2004. We obtained the U.S. Environmental Protection Agency’s 1999 modeled estimates of benzene and 1,3-butadiene for 886 census tracts surrounding Houston, Texas. We ran Poisson regression …

Endoplasmic Reticulum Stress Markers Are Associated With Obesity In Nondiabetic Subjects, Neeraj K. Sharma, Swapan K. Das, Ashis K. Mondal, Oksana Hackney, Winston S. Chu, Philip A. Kern, Neda Rasouli, Horace J. Spencer, Aiwei Yao-Borengasser, Steven C. Elbein

Clinical and Translational Science Faculty Publications

Objective: Adipocyte and hepatocyte endoplasmic reticulum (ER) stress response is activated in dietary and genetic models of obesity in mice. We hypothesized that ER stress was also activated and associated with reduced insulin sensitivity (S_I) in human obesity.

Research Design and Methods: We recruited 78 healthy, nondiabetic individuals over a spectrum of body mass index (BMI) who underwent oral and iv glucose tolerance tests, and fasting sc adipose and muscle biopsies. We tested expression of 18 genes and levels of total and phosphorylated eukaryotic initiation factor 2α, c-jun, and c-Jun N-terminal kinase 1 in adipose tissue. We compared …

Stearoyl-Coenzyme A Desaturase 1 Gene Expression Increases After Pioglitazone Treatment And Is Associated With Peroxisomal Proliferator-Activated Receptor-Γ Responsiveness, Aiwei Yao-Borengasser, Negah Rassouli, Vijayalakshmi Varma, Angela M. Bodles, Neda Rasouli, Resat Unal, Bounleut Phanavanh, Gouri Ranganathan, Robert E. Mcgehee Jr., Philip A. Kern

Clinical and Translational Science Faculty Publications

Context and Objective: Stearoyl-coenzyme A desaturase (SCD1) is the rate-limiting enzyme that converts palmitoyl- and stearoyl-coenzyme A to palmitoleoyl- and oleoyl-cownzyme A, respectively. SCD-deficient mice are protected from obesity, and the ob/ob mouse has high levels of SCD. This study was designed to better characterize SCD1 gene and protein expression in humans with varying insulin sensitivity.

Design, Participants, and Setting: In a university hospital clinical research center setting, SCD1 gene expression was measured in sc adipose and vastus lateralis muscle of 86 nondiabetic subjects; 10 wk of pioglitazone (45 mg daily) and metformin (1000 mg twice daily) treatment were assessed …

Technical Variables In High-Throughput Mirna Expression Profiling: Much Work Remains To Be Done, Peter T. Nelson, Wang-Xia Wang, Bernard R. Wilfred, Guiliang Tang

Pathology and Laboratory Medicine Faculty Publications

MicroRNA (miRNA) gene expression profiling has provided important insights into plant and animal biology. However, there has not been ample published work about pitfalls associated with technical parameters in miRNA gene expression profiling. One source of pertinent information about technical variables in gene expression profiling is the separate and more well-established literature regarding mRNA expression profiling. However, many aspects of miRNA biochemistry are unique. For example, the cellular processing and compartmentation of miRNAs, the differential stability of specific miRNAs, and aspects of global miRNA expression regulation require specific consideration. Additional possible sources of systematic bias in miRNA expression studies include …

Focus On Rna Isolation: Obtaining Rna For Microrna (Mirna) Expression Profiling Analyses Of Neural Tissue, Wang-Xia Wang, Bernard R. Wilfred, Donald A. Baldwin, R. Benjamin Isett, Na Ren, Arnold J. Stromberg, Peter T. Nelson

Sanders-Brown Center on Aging Faculty Publications

MicroRNAs (miRNAs) are present in all known plant and animal tissues and appear to be somewhat concentrated in the mammalian nervous system. Many different miRNA expression profiling platforms have been described. However, relatively little research has been published to establish the importance of 'upstream' variables in RNA isolation for neural miRNA expression profiling. We tested whether apparent changes in miRNA expression profiles may be associated with tissue processing, RNA isolation techniques, or different cell types in the sample. RNA isolation was performed on a single brain sample using eight different RNA isolation methods, and results were correlated using a conventional …

Roofing Foreman Dies From Telescopic Boom Lift Fall, Kentucky Injury Prevention And Research Center

Fatality Case Reports--Falls

On a winter day in 2008, a male roofer/ foreman died after falling from a telescopic boom lift. He and four other roofing crew members were installing a new roof at a residence undergoing restoration. They had arrived at the job site at approximately 7:45 AM.

At approximately 8:00 AM, the foreman and two roofers were installing flashing on the roof of an alcove on the south side of the house while two other roofers were in a pickup truck putting on coveralls. The crew on the roof needed red rosin underlayment (paper) which was in the pickup truck. Using …

Sequiterpene Synthase Gene And Protein, Joe Chappell, Bryan T. Greenhagen

Pharmaceutical Sciences Faculty Patents

The invention relates to sesquiterpene synthases and methods for their production and use. Particularly, the invention provides nucleic acids comprising the nucleotide sequence of citrus valencene synthase (CVS) which codes for at least one CVS. The invention further provides nucleic acids comprising the nucleotide sequence coding for amino acid residues forming the tier 1 and tier 2 domains of CVS. The invention also provides for methods of making and using the nucleic acids and amino acids of the current invention.

Focal Cerebral Ischemia In The Tnfalpha-Transgenic Rat, L. Creed Pettigrew, Mark S. Kindy, Stephen W. Scheff, Joe E. Springer, Richard J. Kryscio, Yizhao Li, David S. Grass

Sanders-Brown Center on Aging Faculty Publications

BACKGROUND: To determine if chronic elevation of the inflammatory cytokine, tumor necrosis factor-alpha (TNFalpha), will affect infarct volume or cortical perfusion after focal cerebral ischemia.

METHODS: Transgenic (TNFalpha-Tg) rats overexpressing the murine TNFalpha gene in brain were prepared by injection of mouse DNA into rat oocytes. Brain levels of TNFalpha mRNA and protein were measured and compared between TNFalpha-Tg and non-transgenic (non-Tg) littermates. Mean infarct volume was calculated 24 hours or 7 days after one hour of reversible middle cerebral artery occlusion (MCAO). Cortical perfusion was monitored by laser-Doppler flowmetry (LDF) during MCAO. Cortical vascular density was quantified by stereology. …

High-Activity Mutants Of Butyrylcholinesterase For Cocaine Hydrolysis And Method Of Generating The Same, Chang-Guo Zhan, Hoon Cho, Hsin-Hsiung Tai

Pharmaceutical Sciences Faculty Patents

A novel computational method and generation of mutant butyrylcholinesterase for cocaine hydrolysis is provided. The method includes molecular modeling a possible BChE mutant and conducting molecular dynamics simulations and hybrid quantum mechanical/molecular mechanical calculations thereby providing a screening method of possible BChE mutants by predicting which mutant will lead to a more stable transition state for a rate determining step. Site-directed mutagenesis, protein expression, and protein activity is conducted for mutants determined computationally as being good candidates for possible BChE mutants, i.e., ones predicted to have higher catalytic efficiency as compared with wild-type BChE. In addition, mutants A199S/A328W/Y332G, A199S/F227A/A328W/Y332G, A199S/S287G/A328W/Y332G, …

Missense Mutation In Exon 2 Of Slc36a1 Responsible For Champagne Dilution In Horses, Deborah Cook, Samantha Brooks, Rebecca Bellone, Ernest Bailey

Veterinary Science Faculty Publications

Champagne coat color in horses is controlled by a single, autosomal-dominant gene (CH). The phenotype produced by this gene is valued by many horse breeders, but can be difficult to distinguish from the effect produced by the Cream coat color dilution gene (CR). Three sires and their families segregating for CH were tested by genome scanning with microsatellite markers. The CH gene was mapped within a 6 cM region on horse chromosome 14 (LOD = 11.74 for theta = 0.00). Four candidate genes were identified within the region, namely SPARC [Secreted protein, acidic, cysteine-rich (osteonectin)], SLC36A1 (Solute Carrier 36 family …

Method For Enhancing Protective Cellular Responses To Genotoxic Stress In Skin, Elaine L. Jacobson, Myron K. Jacobson

Pharmaceutical Sciences Faculty Patents

The present invention is directed to methods of using pro-NAD agents capable of enhancing the dermal and epidermal skin cell NAD content. These pro-NAD agents may be administered topically, orally, or parenterally to enhance DNA repair and other protective responses to DNA damage. The invention further relates to pharmaceutical compositions comprising pro-NAD agents that effectively elevate intracellular NAD content. Finally, the invention relates to the method of using the pro-NAD agents to treat disorders such as sunburn and other skin deterioration that results from DNA damage in skin cells.

Derivatives Of Mithramycin And Methods Of Making And Uses Thereof, Jürgen Rohr, Lily L. Remsing, Mohammad Ner-E-Alam, Jose A. Salas, Carmen Mendez, Alfredo F. Brana, Ana M. Gonzalez

Pharmaceutical Sciences Faculty Patents

The invention, in one aspect, generally relates to mithramycin derivatives from mutated Streptomyces argillaceus and their production. The invention also relates using the derivatives for the treatment of various diseases. Finally, the invention relates to a mutated Streptomyces argillaceus useful in the production of the mithramycin derivatives.

A 53-Year-Old Male Iron Foreman Dies After Fall From Steel Decking, Kentucky Injury Prevention And Research Center

Fatality Case Reports--Falls

During the fall of 2007, a 53-year-old male iron foreman died when he fell approximately 18 feet from roofing being installed on a retail building under new construction. Two work crews were installing metal decking for a roof by laying corrugated metal sheets to the joists of a retail building being constructed. All workers wore personal fall arrest systems complete with lanyards on the roof; none of the workers were tied off to the building. The foreman had accessed the work site via a 32-foot extension ladder to check on the progress of the work. After speaking to the first …

Review Of Oral Oxymorphone In The Management Of Pain, Paul Sloan

Anesthesiology Faculty Publications

Chronic cancer and nonmalignant pain (CNMP) is a common and major health problem afflicting approximately 40 million persons in the US. Most cancer patients, and many patients with CNMP, require opioid analgesics to obtain adequate pain relief. Oral oxymorphone is a new formulation of an existing parenteral opioid that has become available for the treatment of significant pain: acute postoperative, chronic arthritis, chronic low back, and chronic cancer pain. Oxymorphone is a typical mu-opioid agonist that is effective in both immediate- and extended-release (IR and ER) formulations. Oxymorphone is more lipid soluble than morphine, resulting in a rapid onset of …

Accelerated High Fidelity Prion Amplification Within And Across Prion Species Barriers, Kristi M. Green, Joaquín Castilla, Tanya S. Seward, Dana L. Napier, Jean E. Jewell, Claudio Soto, Glenn C. Telling

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Experimental obstacles have impeded our ability to study prion transmission within and, more particularly, between species. Here, we used cervid prion protein expressed in brain extracts of transgenic mice, referred to as Tg(CerPrP), as a substrate for in vitro generation of chronic wasting disease (CWD) prions by protein misfolding cyclic amplification (PMCA). Characterization of this infectivity in Tg(CerPrP) mice demonstrated that serial PMCA resulted in the high fidelity amplification of CWD prions with apparently unaltered properties. Using similar methods to amplify mouse RML prions and characterize the resulting novel cervid prions, we show that serial PMCA abrogated a transmission barrier …