In Vitro and Animal Models for Emerging Diseases and Biodefense,”--Part A: “In Vitro Screens for Antimicrobial Activity”
As directed by the NIH project officer, the Center for Veterinary Health Sciences, Oklahoma State University, has developed, validated, and is currently using in vitro assays to screen test substances for activity against emerging “infectious agents.” The activity in this contract involves the screening of compounds for antimicrobial activity against several biodefense-related select agents in Categories A and B. Beginning October 2, 2006, new task orders will include drug screening against several drug-resistant pathogenic bacteria as well as genetic finger printing of organisms involved in the drug screening process. Also beginning October 2, 2006, the research team will initiate drug screening of a chemical library consisting of 10,000 drug-like compounds for activity against biodefense-related select agents. This information will be useful for research scientists around the country to better evaluate their own chemical libraries for compounds with potential activity against these select agents. This will in turn stimulate the submission of additional compounds into the NIH drug screening program. This is an IDIQ contract that runs through September 29, 2010, with a maximum potential budget of $40 million. Thus far, more than $5 million has been awarded for tasks orders. Materials for testing will be provided by NIAID, as described in RFP NIH-NIAID-DMID-03-39.
Sponsor: NIH, NIAID, DMID, Contract HHSN266200400004I.
PI: W.W. Barrow, K. Clinkenbeard, R. Morton, P. Bourne, M. Wright-Valderas
A Cell Culture-Derived Vaccine for Anaplasmosis
Anaplasma marginale harvested from tick cell culture will be tested as an antigen for a new and improved vaccine for bovine anaplasmosis. This antigen should result in the development of a vaccine that is safe, easily standardized, and free of contaminating bovine cells and pathogens. A vaccine dose will be formulated and tested in cattle for vaccine efficacy.
Sponsors: Novartis Animal Vaccines, Inc., Oklahoma Center for the Advancement of Science and Technology
PIs: Edmour F. Blouin, Katherine M. Kocan, Jose de la Fuenta
Novartis Animal Vaccines, Inc.: Thomas Halbur, Virginia C. Onet
Development of Aptamer Beacons to Lipopolysaccharide for the Real-time Sensing of BW Agents
Force protection is of utmost importance, but a lack of real-time sensing technologies for biological warfare (BW) agents leaves US forces vulnerable to conventional as well as terrorist’s BW attacks. The major barriers to real-time sensing of BW agents are: 1) a lack of robust probes for detection of BW agents, and 2) inadequate sensitivity of sensor platforms through which target probes transduce their detection signals. We propose to conduct a “proof-of-concept” project to marry a new type of probe technology with exceptional robustness termed aptamers with a newly conceived sensing platform with exceptional sensitivity termed AFP beacons to overcome the current barriers for developing real-time BW sensing. Aptamers are highly stable and specific oligonucleotides, which work like monoclonal antibodies (Mab) to bind directly to BW target agents. However, unlike Mab, aptamers have exceptional stability even under field conditions. Aptamer technology is only beginning to be applied to detection of infectious agents. We developed aptamers to surface exposed targets on the prototype BW agent, enterohemorrhagic E coli. We will next demonstrate that these aptamers can be used to specifically detect BW agents. In future experiments, we will engineer our anti-E coli aptamers to act as beacons so that when the aptamer binds to its target it will switch from an “off” to an “on” signal by turning on specific fluorescence directly. No consumable reagents are required. Once DEPSCoR proof-of-concept project is accomplished we will transition to prototype sensor development using this technology.
Sponsor: Army Research Laboratory
PI: Ken Clinkenbeard
Development of Aptamer Beacons for Antemortem Diagnosis of Chronic Wasting Disease
The goal
of the project is to develop the basic research in support of an aptamer beacon
based antemortem diagnostic test for the transmissible spongiform encephalopathy
chronic wasting disease (CWD) of elk and deer. Currently available diagnostic
tests are based on monoclonal antibodies. These tests are applicable for biopsy
or postmortem samples, but may have limitations that preclude their further
development as antemortem tests. The limitations of Mab-based tests may be
overcome by a new molecular entity known as aptamers. In particular, aptamers
can be manipulated using standard molecular biology techniques to act as beacons
or molecular switches that turn "on" a fluorescent signal when they
bind to their target. Aptamers are nucleic acids which serve as novel recognition
molecules or probes that are highly specific for a wide range of targets.
Attempts by others to use aptamers to distinguish normal hamster prion from
its abnormal isoform were not successful. However, a novel aptamer selection
strategy is proposed herein to overcome the problems of the original aptamer
selection experiments. We propose to use a cross-over selection strategy.
Initially, the aptamer selection target will be particular peptides of CWD
prion thought to be exposed on abnormal isoform. The reduced aptamer pool
will subsequently be selected against the insoluble plaques of the abnormal
isoform isolated from brain tissue of affected elk or deer. Aptamer that specifically
recognize the abnormal isoform of CWD prion will subsequently be engineered
as aptamer beacons, a novel signaling aptamer that have a built-in molecular
switch.
Sponsor: U.S.
Army Medical Research and Materials Command, National Prion Research Program
PI: Ken
Clinkenbeard
Polymer-Based Yersinia Pestis Point-of-Case Diagnostics
The goal of the proposed research is to develop a highly sensitive and specific multi-locus array diagnostic for rapid identification of Yersinia pestis, the causative agent of plague. This infectious bacterial agent is on the NIAID category A priority list as a potential biological warfare (BW) agent. In order to confirm suspicions that clinical cases may be due to purposeful aerosol dissemination of Y. pestis and, as such, may be indicative of an imminent plague epidemic, and to enable healthcare professionals to instigate immediate and effective therapeutic intervention and control measures, a rapid, sensitive, and accurate method for early detection of disease is a high priority. To meet this diagnostic challenge, the detection technology will use a polymer that exhibits intrinsic amplification of fluorescence transduction events to rapidly identify species-specific genomic, proteomic, and lipo-oligosaccharide (LOS) markers of Y. pestis. This amplifying fluorescent polymer (AFP) will be fabricated as nanoparticles and functionalized for covalent attachment of quencher-labeled molecular or aptamer beacon probes, which will trigger amplified fluorescent responses when binding of the target to the probe causes dequenching of the polymer. We are confident that the amplification afforded by AFP will enable detection of target analytes in extremely low concentrations with minimal sample preparation, thus providing significant advantages over current microbiological and molecular diagnostic methods. Once proof of concept is established using standard microarray technology, the diagnostic platform will be integrated into a real-time, low-density, multi-locus array printed in a membrane sample delivery system that ultimately will be used to identify and discriminate between many different BW pathogens.
Sponsors: National Institute for Allergies and Infectious Disease, Nomadics, Inc.
PI: Ken Clinkenbeard, Jerry Malayer
Ruminant B-Lymphocyte Yellow Fluorescent Protein Aggregation Bioassay for Elk Chronic Wasting Disease
The goal of the proposed research is to develop a cell culture model for elk chronic wasting disease (CWD) prion propagation that can be used as a bioassay for detecting CWD. CWD is a transmissible spongiform encephalopathy (TSE), caused by the mis-folding of a normal cell surface prion protein (PrPc) through the interaction with infectious mis-folded and protease resistant prion protein (PrPres). The PrPres specific to CWD is PrPcwd. CWD occurs in free ranging and captive elk and deer herds in several Rocky Mountain and Plains states. Although there is evidence suggesting that CWD cannot be transmitted to humans, this potential has not been thoroughly ruled out. Current infectivity bioassays involve the use of live animals, whereas the proposed cell culture model will reduce the need for animal experimentation to study the mechanism of prion infectivity and disease. In addition, a cell culture will be developed as an assay for the screening of infectious prions in veterinary medical samples from elk and deer. This assay system also has the potential for use in assessing therapeutic strategies. Our goal will be accomplished by bioengineering a bovine B-lymphocyte cell (B-cell) line to surface express elk PrPc fused to yellow fluorescent protein (YFP). Like other glycosyl-phosphoinositol (GPI) anchored proteins, surface expressed YFP-PrPc will have a dispersed distribution on these B-cells. Interaction of these B-cells with infectious, mis-folded, protease resistant CWD prion (PrPcwd) will induce conversion of the dispersed YFP-PrPc to aggregated mis-folded YFP-PrPcwd that will be detected by confocal microscopy as aggregated YFP-PrPcwd on the B-cell surface.
Sponsors: US Army Research Office, Nomadics, Inc.
PI: Ken Clinkenbeard, Jeff Blair
Shipping Fever: New Approaches to Understanding Prevention and Management
The objective is to determine infectious agents and host responses
that cause respiratory disease and/or defend cattle from disease. Naturally
occurring cattle diseases at Noble Foundation, privately owned, USDA, and
OSU herds are being investigated. Emphasis is primarily on bovine viral diarrhea
virus infection and pasteurellosis. Vaccination programs are being evaluated
and new recommendations given.
Sponsor: The Noble Foundation
PIs: Anthony W. Confer, S. Mady Dabo, Robert W. Fulton, Jerry W. Ritchey
Department of Veterinary Clinical Sciences: John G. Kirkpatrick, Robert A.
Smith
The project determines changing patterns, geographical differences, risk factors, and management practices related to bovine respiratory disease. The influence of various bacteria and viruses is studied. In addition, the pharmacokinetics and efficacy of newer therapies and new-generation vaccines are evaluated. The host-pathogen relationship is characterized at the molecular level.
Sponsor: Oklahoma Agricultural Experiment Station
PIs: A. W. Confer, R. W. Fulton, R. J. Panciera, K. D. Clinkenbeard, R. J. Morton
Mannheimia haemolytica Outer Membrane Protein PlpE: Characterization of Epitopes Stimulating Homologous and Heterologous Serotype Protection
This project is a molecular and immunologic approach to studying an immunologically important outer membrane protein of M. haemolytica. It compares the PlpE protein from serotypes 1, 2, and 6 with respect to important epitopes for immunity.
Sponsor: USDA CSREES, National Research Initiative Competitive Grant
PIs: A. W. Confer, Sahlu Ayalew
Mannheimia haemolytica Bacterin-Toxoid Efficacy Studies
This project studies the efficacy of commercial M. haemolytica vaccines in an experimental challenge model.
Sponsor: Pfizer Animal Health
PIs: A. W. Confer, R. J. Panciera
Chimeric Vaccine for Mannheimia haemolytica Infection in Cattle
Recombinant DNA technology is being applied to induce a single chimeric protein that will stimulate immunity in cattle to M. haemolytica leukotoxin and the outer membrane.
Sponsor: Oklahoma Applied Research Program, Oklahoma Center for the Advancement of Science and Technology (OCAST)
PIs: A. W. Confer, Sahlu Ayalew
Pasteurella multocida OmpA: Functional Characterization
This project studies the adherence and colonization properties of Pasteurella multocida OmpA. It is designed to investigate the specific role of P. multocida OmpA in the bacterium interaction with host cells and in the pathogenesis of the disease.
Sponsor: USDA CSREES, National Research Initiative Competitive Grant
PIs: S. M. Dabo, A.W. Confer
Development of a Vaccine Against Ixodes scapularis Infestations
Antigens identified in previous studies to induce protection against Ixodes scapularis tick infestations in mice will be used in vaccine formulations. The vaccine will be tested against the three tick stages: larvae, nymphs, and adults.
Sponsor: Pfizer Animal Health, Inc.
PIs: José de la Fuente, Katherine M. Kocan, Edmour F. Blouin, Consuelo Almazán
The major danger to research and animal care personnel working with rhesus monkeys is monkey B virus (BV), a herpesvirus that is transmitted by bites and scratches and is rapidly fatal in humans if not detected and treated rapidly. This project involves molecular characterization of BV and related viruses of other primates and application of these data to develop more sensitive and specific diagnostic tests that will permit rapid and reliable identification of BV infections in humans.
Sponsor: NIH, NCRR
PI: R. Eberle
Baboons are an important animal species used in biomedical research. This program will develop a breeding colony of baboons in Oklahoma and supports research aimed at improving the breeding efficiency of baboons in captivity, defining viruses that naturally infect baboons, and improving the basic well-being and behavior of captive-bred baboons.
Sponsor: NIH, NCRR
PIs: R. Eberle, A. Kocan, J. d'Offay
OUHSC: Gary White
Indigenous viruses can have a major adverse effect on the results of biomedical research studies using animals, particularly where immunosuppression is involved. This program supports derivation of a colony of baboons that are free of all known herpesviruses and most retroviruses.
Sponsor: NIH, NCRR
PI: R. Eberle
OUHSC: Gary White
Genetic Basis of Drug Resistant Mutants of B Virus
Monkey B virus is a BSL-4 virus that is extremely neurovirulent when transmitted from macaques to humans. Spontaneous mutants of B virus have been isolated that are completely resistant to antiviral drugs used to treat infected humans. The two viral genes that confer drug resistance in the related human virus, herpes simplex virus, do not appear to be involved in drug resistance in B virus. This project will identify the viral genes responsible for drug resistance in B virus.
Sponsor: NIH, NIAID
PI: R. Eberle
Bovine Viral Diarrhea Disease Virus (BVDV) Vaccines: Antibody Response to Heterologous BVDV Strains
The study will determine the range of heterologous immunity in calves receiving modified live virus (MLV) or killed BVDV vaccines. Currently there are two recognized antigenic types of BVDV, Type 1 and 2. The study will determine if these vaccines induce antibodies to various Type 1 and 2 viruses.
Sponsors: Grand Laboratories, Inc., Pfizer Animal Health.
PIs: Robert W. Fulton, Anthony W. Confer
Genetic and Antigenic Variability of BVDV in Cattle Infections
Bovine viral diarrhea viruses (BVDV) isolates from the Oklahoma Animal Disease Diagnostic laboratory (OADDL) will be obtained from clinical/necropsy cases. The viruses will be typed as BVDV 1a, 1b, or 2. Potentially there will be additional typing and/or groups. Field isolates from naturally occurring disease, including persistently infected (PI) cattle, will be compared with vaccinal stains and standard reference strains. A phylogenetic survey of the BVDV subtypes from the field isolates, vaccinal strains, and reference strains will be performed to detect relationships among the virus and their genetic stability. Neutralization tests will be performed to compare the subtypes to the vaccinal strains. Potentially new subtypes may warrant additional subtypes in the vaccines.
Sponsor: Oklahoma Agricultural Experiment Station
PIs: Robert W. Fulton, A. W. Confer
Oklahoma Animal Disease Diagnostic Laboratory: J.T. Saliki
Antibiotic Administration and Vaccination with Live Bacterial Vaccine in Calves
This study will determine if an antibiotic given calves that have been administered avirulent Pasteurella haemolytica and Pasteurella multocida vaccine will decrease the immune responses to the immunogens. Calves will receive Micotil antibiotic and Once PMH Pasteurella haemolytica and P. multocida vaccine. The calves’ sera will be tested for P. haemolytica and P. multocida antibodies.
Sponsor: ELANCO Animal Health, Division of Eli Lilly and Company
PIs: Robert W. Fulton, Anthony W. Confer
Bovine viral diarrhea viruses (BVDV) occur as biotypes, cytopathic (CP) and noncytopathic (NCP), and as genotypes, 1 and 2. Certain BVDV disease forms occur with different biotypes/genotypes. The molecular differences among biotypes/genotypes will be investigated by PCR and nucleic acid sequencing. Virulence markers of BVDV will be investigated.
Sponsor: Oklahoma Agricultural Experiment Station
PIs: Robert W. Fulton, Jean M. d’Offay, Anthony W. Confer, Jerry W. Ritchey
Oklahoma Animal Disease Diagnostic Laboratory: Jeremiah T. Saliki
Evaluation of Viral Vaccine Containing Infectious Bovine Rhinotracheitis Virus (IBRV), Bovine Viral Diarrhea Virus 1 and 2 (BVDV), Parainfluenza -3V (PI-3V), and Bovine Respiratory Syncytial Virus (BRSV) in Preventing Infection and Respiratory Disease in Cattle
The purpose of the study will be to determine if pre-weaning vaccination of ranch calves with viral vaccine: 1) reduces respiratory disease, and 2) reduces transmission of viruses in calves moved from auction markets and commingled with the fresh calves under feedlot conditions.
Sponsor: Fort Dodge Animal Health
PIs: Robert Fulton, A.W. Confer
Oklahoma Animal Disease Diagnostic Laboratory: J.T. Saliki
Department of Veterinary Clinical Sciences: D.L. Step
Vaccination of Ranch Calves with Modified Live Viral Vaccine: Effects on Viral Transmission on Commingled and Transported Calves
This study will determine if vaccination of ranch calves with a modified live viral vaccine will reduce respiratory disease and reduce transmission of viruses in the calves commingled with auction calves and moved to a feedlot.
Sponsor: Schering Plough Animal Health Corp.
PIs: Robert W. Fulton, Anthony W. Confer
Food Safety: Farm to Table
The long-term objectives of this study are to develop methods for assuring the microbial safety of the food supply from farm to table. Focus is on control of Salmonella species and Escherichia coli O157:H7 associated with swine and cattle.
Sponsor: USDA/CSREES
PIs: Terry Lehenbauer
College of Agricultural Sciences and Natural Resources: Stanley Gilliland, Guolong Zhang, Peter Muriana, Siobhan Reilly
Natural History of Borrelia lonestari
This project examines the natural maintenance cycle of Borrelia lonestari, a putative agent of southern tick-associated rash illness (STARI) or “southern Lyme disease” in lone star ticks, white-tailed deer, and other wildlife species. The work involves both evaluation of naturally infected ticks and wild animals, and experimental confirmation of the proposed maintenance cycle.
Sponsor: NIH NIAID
PIs: Susan E. Little, Edmour Blouin, Kathy Kocan
UGA: Michael Yabsley, Kevin Keel
Diagnosis of Borrelia lonestari
This project seeks to develop better diagnostic assays for Borrelia lonestari, a putative agent of southern tick-associated rash illness (STARI) or “southern Lyme disease.” Research projects include experimental infection of white-tailed deer and rabbits and development of microbiologic, molecular, and serologic techniques to identify infected animals with the ultimate goal of developing diagnostic assays for use in people.
Sponsor: NIH NIAID
PIs: Susan E. Little
UGA: Michael Yabsley
Transmission of Ehrlichia canis by Rhipicephalus sanguineus
This project examines the transmission dynamics at play as Ehrlichia canis is moved between dogs by the brown dog tick Rhipicephalus sanguineus.
Sponsor: Bayer Animal Health.
PIs: Susan E. Little, Kathy Kocan, Eileen Johnson, Sidney Ewing
Ehrlichia ewingii Infection and Exposure Rates in Dogs
Ehrlichia ewingii commonly infects dogs in areas of the US where lone star ticks predominate. This project seeks to document the prevalence of E. ewingii infection and exposure in dogs from the Ozark Plateau and use samples acquired from naturally infected dogs to refine diagnostic assays for E. ewingii and other, closely-related rickettsial pathogens of dogs and people.
Sponsor: IDEXX
PI: Susan E. Little, Sidney Ewing, Jim Meinkoth
Revealing the Attenuating Mutations of F. tularensis LVS
Francisella tularensis, the causative agent of tularemia, is a highly infectious agent that causes severe disease in mammals, including humans. As such, it is classified as a Category A biodefense agent. The virulence mechanisms for this agent have not been elucidated. An attenuated strain of F. tularensis (LVS or Live Vaccine Strain), which has shown to be effective against human tularemia has not been licensed in this country, mainly because it is a live vaccine and the cause for its attenuation is unknown. The close genomic relationship (99.9% identity) between the LVS and a wild type strain (OSU-18) allows the identification, categorization, and ranking of all the genetic differences for their attenuating potential. From this analysis, complemented LVS and knockout OSU18 mutant strains will be tested in a murine model of infection, which should identify genes involved in attenuation. Such information also will be valuable in constructing additional attenuated strains that may be used for vaccines as well as revealing important virulence factors for this agent.
Sponsor: Western Regional Center for Biodefense and Emerging Diseases
PIs: Rebecca J. Morton, Ken Clinkenbeard
Baylor College of Medicine: Joseph Petrosino, George Weinstock
Comparative Study of Experimental Francisella tularensis Type A and B in Cats
Tularemia, caused by Francisella tularensis, is a zoonotic disease observed mainly in mammalian wildlife. The disease in humans and animals can be severe and may result in death. The disease in the United States is complicated by the presence of two major subspecies that differ in virulence and ecology. Another complicating factor is the well-documented potential of F. tularensis as an agent of bioterrorism. Because the disease is being recognized with increasing frequency in cats, information on the clinical and diagnostic parameters of feline tularemia is needed to enable veterinarians to diagnose the disease accurately and rapidly. This project is designed to compare the clinical signs, clinical laboratory parameters, and pathology of experimental tularemia in cats infected with either F. tularensis subsp. tularensis (Type A) or F. tularensis subsp. holarctica (Type B).
Sponsor: OSU Center for Veterinary Health Sciences
PIs: Rebecca Morton, Melanie Breshears, Roger Panciera, Jim Meinkoth, Mason Reichard
Cytokine Expression in Response to Brucella Vaccines
Cytokine responses induced by Brucella polysaccharides in blood cells from vaccinated and unvaccinated cattle and mice, unvaccinated pigs, and humans were evaluated in vitro. Reverse transcriptase-polymerase chain reaction was used to detect cytokine mRNA. Development of a cytokine diagnostic profile indicative of induction of immunity for application to human vaccinates was attempted.
Sponsor: Canada Department of National Defense
PI: John H. Wyckoff III
Synthetic peptides with sequences homologous to Mycobacterium bovis proteins will be used to develop better test reagents for field and laboratory diagnosis of bovine tuberculosis. Cattle immunized with M. bovis will be compared for responsiveness to these antigens and those of M. avium to determine both specific and cross-reactive responses.
Sponsor: Oklahoma Agricultural Experiment Station
PIs: John H. Wyckoff III
Heat shock protein-specific T lymphocytes derived from Brucella abortus vaccinated cattle will be characterized by flow cytometry for surface marker expression and cytotoxic function against infected monocyte-derived macrophages. Cytokine production by the T lymphocytes will be analyzed through RT-PCR. These studies will define a host defense effector mechanism against brucellosis.
Sponsor: United States Department of Agriculture
PIs: John H. Wyckoff III, Anthony W. Confer
DEPARTMENT OF PHYSIOLOGICAL SCIENCES
Trans-Differentiation of Alveolar Type II Epithelial Cells to Type I: Role of TGF β1 Pathway
The goal of this student seed grant is to study the role of TGF in the differentiation of alveolar epithelial cells.
Sponsor: Center of Veterinary Health Sciences, Oklahoma State University
PI: Manoj Bhaskaran (Mentor: Lin Liu)
The major goals of this project are to elucidate human sulfotransferase (SULT) chemical and kinetic mechanisms, to understand physiologic functions of SULTs, and to investigate their relevance to human health in physiologic and pathologic conditions. Research focus on: 1) mechanisms of enzyme catalysis, substrate inhibition, and product activation of human SULTs; 2) effect of clinical, widely used drugs on human sulfotransferase catalytic activities; and 3) oxidative regulation mechanisms of human SULTs.
Sponsor: National Institute of Health
PI: Guangping Chen
Division of Agricultural Sciences and Natural Resources: Richard Essenberg
University of Oklahoma: Paul Cook
Bioactive Food Components and Human Sulfotransferases
Many bioactive food components and active components in drugs are phenolics. These phenolics are substrates for human sulfotransferases (SULTs). They also inhibit SULTs (substrate-inhibition). Our recent data suggests that these compounds also induce human SULTs. This research project focuses on the relationships between bioactive food components and human SULTs, including how they are metabolized by SULTs, how they inhibit SULTs, and how they regulate human SULT genes.
Sponsor: United States Department of Agriculture
PI: Guangping Chen
Studies in this project focus on sulfotransferase (SULT) induction by methotrexate. Rats, human hepatic carcinoma cell line, Hep G2, and human intestinal carcinoma cell line, Caco-2, will be used for these studies. Enzyme activity assay, Western blot, RT-PCR, site directed mutagenesis, plasmid transfection, small interfering RNA (siRNA) gene silencing, promoter gene deletion, DNA footprinting, and electrophoretic mobility shift assay will be used to determine the SULT gene regulation and nuclear receptor mediated SULT induction mechanisms.
Sponsors: Oklahoma Center for the Advancement of Science and Technology, Oklahoma State University
PI: Guangping Chen
Proteomic Analysis of Lipid Rafts Isolated from Alveolar Type II Cells
The goal of this student seed grant is to identify protein components in lipid rafts of alveolar type II cells using a proteomic approach.
Sponsor: Center of Veterinary Health Sciences, Oklahoma State University
PI: Narendranath Reddy Chintagari (Mentor: Lin Liu)
Lipid Rafts: Implications for Surfactant Secretion by Alveolar Type II Cells
The goal of this student seed grant is to study the role of lipid rafts in lung surfactant secretion.
Sponsor: Center of Veterinary Health Sciences, Oklahoma State University
PI: Narendranath Reddy Chintagari (Mentor: Lin Liu)
The long term goal of the project is to evaluate the contribution of the mar regulon to survival of Salmonella serovar Typhimurium in vivo, in the absence of antibiotics or in the presence of subtherapeutic, and the therapeutic levels of antibiotics.
Sponsor: OCAST Health Research
PI: Cyril Clarke
The objective of this research is to develop a novel detection system utilizing an amplifying fluorescent polymer for rapid, highly sensitive, and selective detection of methicillin-resistant Staphylococcus aureus present in clinical exudates without prior culture and isolation of bacteria.
Sponsors: Nomadics, Inc, National Institutes of Health
PIs: Cyril Clarke, Jerry R. Malayer
Molecular
Mechanisms of Lung Surfactant Secretion
Lung surfactant is a surface-active material that stabilizes alveoli. It is
synthesized and secreted by lung epithelial type II cells. The long-term
objective of this proposed project is to elucidate the molecular mechanisms of
lung surfactant secretion from type II cells including transport of secretory
granules to and fusion with the plasma membrane. Deficiency of lung surfactant
is the cause of respiratory distress syndrome in premature infants.
Accomplishing the goals of this proposal may give a valuable insight to the
therapy of pulmonary diseases such as neonatal respiratory distress syndrome.
Sponsor: National Institutes of Health
PI: Lin Liu
Mechanisms of Alveolar Epithelial Cell Differentiation
The goal of this grant is to understand molecular mechanisms of the differentiation of alveolar epithelial cells in isolated alveolar epithelial cells, fetal lung development, repair of injured lungs using in-house made 10K rat gene arrays, and RNA interference.
Sponsor: National Institutes of Health
PI: Lin Liu
Mechanisms of Alveolar Fluid Transport
The goal of this project is to investigate the roles of chloride channels of alveolar epithelial type I and type II cells in fluid secretion of fetal lungs, and in maintaining fluid homeostasis of adult and injured lungs.
Sponsor: National Institutes of Health
PI: Lin Liu
GABA Receptor and Pulmonary Fluid Transport
The major goal of this project is to identify GABA receptor in type II cells as it relates to chloride secretion.
Sponsor: March of Dimes Birth Defects
PI: Lin Liu
Biotechnology, Gene Silencing Using RNA Interference
The goal of this project is to develop gene silencing technology.
Sponsor: Oklahoma State Reagents of Higher Education
PI: Lin Liu
Systemic Inflammation and α-synuclein Overexpression are Predisposing Factors in Development of Spontaneous Dopaminergic Neurodegenerative Disease
Accumulation and misfolding of the nerve protein α-synuclein is a hallmark of Parkinson’s disease, a dopaminergic neurodegenerative condition. The events that initiate α-synuclein accumulation and development of neurodegeneration in spontaneous disease are unknown. This project investigates the role of inflammation in development of a dopaminergic neurodegenerative disease of horses, equine pituitary pars intermedia dysfunction (PPID). Specifically, this project will determine if expression of 1) α-synuclein, and 2) inflammatory cytokines is greater following inflammatory stimulation of white blood cells from horses with PPID compared to healthy horses.
Sponsor: Research Advisory Committee, CVHS
PIs: Dianne McFarlane
Clinical Sciences: Todd C. Holbrook
Response of Healthy Horses and Horses with Pituitary Pars Intermedia Dysfunction (PPID) to Administration of α-melanocyte Stimulating Hormone
PPID is a common disease of geriatric horses that adversely impacts their quality of life. Recent research suggests that existing diagnostic tests for PPID are not consistently reliable. The goal of this project is to develop a novel, reliable diagnostic test for PPID. PPID results in overproduction of the pituitary hormones, α-MSH and CLIP. We hypothesize that α-MSH and CLIP control their own release via negative feedback in healthy horses, but this feedback is lost in horses with PPID. α-MSH or CLIP will be administered to healthy and PPID horses and plasma α-MSH measured to determine if negative feedback occurs.
Sponsor: Research Advisory Committee, CVHS
PIs: Dianne McFarlane
Clinical Sciences: Peggy Brosnahan
Gene Expression Changes Following Organophosphate Exposure: Effects of Atropine or Cannabinoid as Antidote
Organophosphate intoxication is primarily treated with the antimuscarinic drug atropine. Epidemiological studies suggest persons treated for organophosphate intoxication may have persistent neurological deficits. While atropine is therapeutically beneficial, its ability to increase acetylcholine release in the brain may contribute to persistent neurologic sequelae. We recently determined that cannabinoids may be beneficial in the treatment of organophosphate intoxication. This project will begin evaluation of possible genetic changes that may lead to persistent neurological deficits following organophosphate intoxication, and whether antidotal therapy may differentially alter gene expression.
Sponsor: Center of Veterinary Health Sciences, Oklahoma State University
PI: Anuradha Nallapaneni (Mentor: Carey Pope)
Presynaptic Modulation of Anticholinesterase Toxicity
The project evaluates contribution of presynaptic neurochemical mechanisms (e.g., in particular neurotransmitter release) in differential toxicity of organophosphorus insecticides.
Sponsors: National Institute of Environmental Health Sciences/National Institute of Aging
PIs: Carey Pope, Jing Liu Pope, Guangping Chen
Water-soluble Nanoparticles for Oxygen Imaging in Tumor
Researchers from the Center for Veterinary Health Sciences and Nomadics, Inc. are evaluating the ability of nanomaterials to enhance tumor imaging. Both applications and toxic potential are jointly being investigated.
Sponsors: National Cancer Institute, Nomadics, Inc.
PIs: Carey Pope, Guangping Chen
Nanoparticle Self-lighting Photodynamic Therapy for Ovarian Cancer Treatment
Collaborative studies between researchers at the Center for Veterinary Health Sciences and Nomadics, Inc. are evaluating the potential for nanoparticles coupled to radical generators to be used in photodynamic therapy for cancer. Feasibility and toxicity studies are concurrently conducted.
Sponsors: U.S. Army, Nomadics, Inc.
PIs: Carey Pope, Jing Liu Pope
Scintillation Luminescence System for In Vivo Dosimetry
This collaborative project between investigators at the Center for Veterinary Health Sciences and Nomadics, Inc. evaluates the potential for nanoparticles to be used in measuring radiation dosing in vivo. Feasibility and toxicity studies are concurrently conducted.
Sponsors: National Cancer Institute, Nomadics, Inc.
PIs: Carey Pope, Guangping Chen, Jerry Ritchey, Cyril Clarke
Medical Countermeasures to Chemical Terrorism
This supplemental project for grant 2R01 ES009119 extends evaluation of the comparative effects of anticholinesterases on brain acetylcholinesterase activity and acetylcholine release.
Sponsors: National Institute of Neurological Disorders and Stroke/National Institute of Environmental Health Sciences
PI: Carey Pope
10th Meeting, International Neurotoxicology Association
This project supported the development and conduct of an international scientific meeting in Porvoo, Finland.
Sponsors: National Institute of Neurological Disorders and Stroke/National Institute of Environmental Health Sciences
PI: Carey Pope
Arsenic, Cancer, and Animal Health and Welfare
The goal of this Veterinary Student Fellowship is to study the relationship between arsenic exposure and cancer.
Sponsor: Morris Animal Foundation
PI: Letitia Posey (Mentor: Lin Liu)
Gene Expression Changes Following Low-level Chlorpyrifos Exposure: Implications for Developmental Neurotoxicity
The organophosphate insecticide chlorpyrifos elicits acute toxicity by inhibiting the enzyme acetylcholinesterase. A number of studies suggest that chlorpyrifos can disrupt early development of the mammalian brain independent of acetylcholinesterase inhibition. These studies will evaluate gene expression in brains following early postnatal exposure to chlorpyrifos at sub-toxic dosages.
Sponsor: Center of Veterinary Health Sciences, Oklahoma State University
PI: Anamika Ray (Mentor: Carey Pope)
Functions of miRNAs in Fetal Lung Development
The goal of this student seed grant is to identify microRNAs that are important for fetal lung development using microRNA microarray.
Sponsor: Center of Veterinary Health Sciences, Oklahoma State University
PI: Yang Wang (Mentor: Lin Liu)
Identification of Proteins in Lamellar Bodies Involved in Lung Surfactant
The goal of this student seed grant is to identify protein components in lamellar bodies of lung type II cells using a proteomic approach.
Sponsor: Center of Veterinary Health Sciences, Oklahoma State University
PI: Pengcheng Wang (Mentor: Lin Liu)
Gene Profiling of Rat Fetal Lung Development Using 10K DNA Microarray
The goal of this student seed grant is to identify the genes that are essential for fetal lung development using DNA microarray.
Sponsor: Center of Veterinary Health Sciences, Oklahoma State University
PI: Tingting Weng (Mentor: Lin Liu)
Functional Studies of PTN in Fetal Lung Development
The goal of this predoctoral fellowship is to study roles of PTN in fetal lung development.
Sponor: American Heart Association
PI: Tingting Weng (Mentor: Lin Liu)
Efficacy of Famotidine for the Prevention of Exercise-Induced Gastritis in Racing Alaskan Sled Dogs
Exercise-induced gastritis and gastric ulcers are common in humans, horses, and racing sled dogs. We tested the hypothesis that famotidine would reduce the incidence and severity of exercise-induced gastric disease in sled dogs. A team of 16 fit sled dogs participated. Study results showed that treatment with famotidine significantly reduced gastric lesion severity score compared to control. There was also a strong trend towards the ability of famotidine to reduce the overall prevalence of exercise-induced gastritis. Our data suggest that famotidine is effective in reducing the severity and possibly the prevalence of exercise induced gastric disease in sled dogs.
Sponsor: Comparative Exercise Physiology Laboratory, OSU
PIs: Kathy Williamson, Erica C. McKenzie, Christopher M. Royer, Michael S. Davis
Texas A&M University: M.D. Willard
DEPARTMENT OF VETINARY CLINICAL SCIENCES
Effects of 5% Lidocaine Transdermal Patch on Chronic Equine Distal Limb Lameness
Lidocaine transdermal patches are utilized frequently in humans to help control localized pain. In this study we used 5% lidocaine transdermal patches on naturally occurring cases of lameness in the horse.
Sponsors: Oxley Chair, Equine Sports Medicine
PIs: Dustin Devine, H. David Moll, Ron Erkert
The Effects of Intravenous Lidocaine of Expired Sevoflurane Concentration in Horses Undergoing Surgery
Intravenous lidocaine has been shown to decrease the concentration of isoflurane needed to anesthetize horses. In this study we postulated that it would have similar effects when used with sevoflurane. This could be important especially in athletic horses undergoing surgery.
Sponsers: Oxley Chair, Equine Sports Medicine
PIs: John Marshall, Jeff Ko, H. David Moll
Effects of Chlorhexidine Gluconate of Varying Concentrations on the Histology of Muccosal, Nervous, and Vascular Tissue of the Equine Guttural Pouch
Guttural pouch infection is a common condition causing loss of performance in the horse. In this study we tried different concentrations of chlohexidine gluconate to determine which would be bacteriocidal without harming components of the guttural pouch.
Sponser: Oxley Chair, Equine Sports Medicine
PIs: Todd Holbrook, H. David Moll, Margaret Brosnahan
Effects of Freeze Thaw Cycles on Components of Equine Plasma
Equine plasma is used for replacement on adult horses loosing protein and on foals suffering from septicemia or born with low IgG levels. Plasma is stored frozen. We subjected equine plasma to freeze thaw cycles to determine if any detrimental factors occurred.
Sponser: Oxley Chair, Equine Sports Medicine
PIs:Todd Holbrook, H. David Moll
Evaluation of the In Vivo use of Lidocaine to Attenuate Ischemia-Reperfusion Injury in the Equine Jejunem
An abdominal crisis with strangulation of intestine is a major cause of death in the horse. In this study we utilized lidocaine to establish if it would attenuate the damage done with a strangulating obstruction.
Sponsers: Oxley Chair, Equine Sports Medicine, Morris Animal Foundation, American Quarter Horse Association
PIs: John Marshall, H. David Moll
North Carolina State University: Anthony Bilkslager
Since the establishment of the Biomedical Laser Laboratory within the College of Veterinary Medicine, research to establish protocols for clinical applications in veterinary medicine has been a primary objective. In addition, the use of laboratory models has resulted in transfer of technology to both industry and human medicine. Work will continue concentrating on the clinical applications of biomedical lasers coupled with collaborative research protocols involving basic scientists (engineers, physicists) and clinicians.
Sponsor: McCasland Foundation and the Mercy Works Foundation
PI: Kenneth E. Bartels
Photodynamic Therapy in Combination with Chemotherapy and Immunotherapy for Treatment of Metastatic Mammary Tumors
Photodynamic therapy (PDT) can be an effective means for direct tumor destruction through its selective photochemical reaction. Common sensitizers used in PDT are 5-aminolevulinic acid (ALA) and Photofrin®. By itself, PDT using ALA and Photofrin® can be effective in some cases, but not effective in others. To address this inconsistency, combinations of several drugs can result in synergistic or additive effects for certain types of cancer such as mammary carcinoma. Combining multiple tumor treatment modalities may create a type of “cocktail therapy” that could be successful for primary and metastatic disease caused by some types of cancer in both animals and human beings. It is hypothesized that the combination use of PDT and the glycated chitosan (GC) immunoadjuvant may significantly improve the efficacy of the treatment, particularly in treating metastatic tumors. It is further hypothesized that the concurrent use of PDT and a new chemotherapeutic agent, AEADA (Bis-1, 4 aminoethylamino-5, 8-dihydroxyanthraquinone2HCl), may enhance the systemic effect of cancer treatment. Combinations of these treatment modalities will be evaluated in the rat and mouse mammary tumor models.
Sponsors: Kleberg Foundation, Barbour Foundation
PIs: Kenneth E. Bartels, Wei Chen (University of Central Oklahoma)
Ethiopia Sheep and Goat Productivity Improvement Program
The goal of this study is to increase the productivity of small ruminants in Ethiopia to improve food and economic securities. The three principal objectives of the program are: 1) Improve small ruminant production and marketing practices; 2) determine the preferred means of utilization of indigenous sheep and goats in future production systems; and 3) enhance the communication capacity at collaborating Ethiopian institutions to facilitate information exchange and accessibility, and research collaborative capabilities and distance education capabilities.
Sponsor: USAID Ethiopia
PI: Lionel J. Dawson
Al-Sharaka, The Partnership, Revitalizing the Higher Education System in Iraq
The goal is to revitalize the higher education system in Iraq through training and assistance. Langston University’s role will be in updating the skills of Iraqi scientists in small ruminant production and provision of supplies and training to establish a ruminant rutrition laboratory at Salahaddin University.
Sponsor: USAID
PI: Lionel J. Dawson
Evaluating and Modeling Extended Lactations in Dairy Goats
A major obstacle to profitability of dairy goat is the highly seasonal supply of milk. Some dairy goat producers are extending the lactation period of their does in order to produce milk during the seasonal period of low milk supply, thereby taking advantage of increased commodity prices. Understanding the biological processes operating during extended lactation in goats will provide tools for better management and will increase our understanding of mammary gland biology.
Sponsor: E. (Kika) de la Garza American Institute for Goat Research, Langston University; Animal and National Resource Institute; Clemson University; Oklahoma State University
PI: Lionel J. Dawson
Nutrient Requirements of Goats: Composition of Tissue Gain and Loss by Mature Goats
The overall objective of the study was to determine the composition of tissue loss and gain by mature goats that varied in initial body weight (BW) and body condition score (BCS). The initial BW and BCS were achieved by subjecting mature Boar x Spanish goats to a management scheme that elicited two levels (thin and fat) of body constitution or fatness, prior to the on-set of the experiment. A secondary objective was aimed at developing equations that will enable the prediction of body composition of mature meat goats from shrunk body weight after urea dilution, and the third was to characterize the effects of body condition and nutritional plane on energy metabolism.
Sponsors: College of Veterinary Medicine, Langston University, USDA
PIs: Lionel J. Dawson, A.T. Ngwa, A.L. Goetsch, R. Puchala, G. Detweiller, T. Sahlu, R. Merkel
Enhanced Goat Production Systems for the Southern United States
Overall project goals are to improve the compatibility of goat production systems with available resources throughout the U.S. Supporting objectives are: 1) develop a vehicle to appraise compatibility of available resources and production conditions with goat production systems; 2) project most appropriate goat production systems based on compatibility with presently available resources and production conditions, and evaluate changes in resources or production conditions necessary for employment of alternative, preferred systems; and 3) disseminate and provide training in use of the developed decision-support vehicle.
Sponsor: College of Veterinary Medicine, Langston University and USDA
Co PIs: Lionel J. Dawson, A.L. Goetsch, T. Sahlu, R. Merkel, T. Gipson, S.D. Hart, H. Blackburn, S. Wildeus
Effects of Ginger and Garlic on Nematode Infection in Goats
Gastrointestinal tract parasitism causes a significant economic loss in small ruminant production. Current management of the internal 2parasite problem relies on utilization of anthelmintic drugs. However, this practice encounters a compelling challenge because of widespread parasitic resistance to anthelmintics and the increasing demand for “green” animal products in the consumer market. Therefore, the importance of alternative approaches for parasite control for sustainable small ruminant production can never be overstressed. One of the promising approaches is to enhance protective immunity of host animals.
Sponsor: Langston University, Langston, OK
PIs: Z. Wang, A. L. Goetsch, S. Hart, E. Loetz, T. Sahlu, L. J. Dawson
The Effect of Dietary Protein on Tissue GSH Concentration and Cytokine Expression in Goats
Protein supplementation enhances resistance to internal parasites in animals. Compared with those fed a straw-based diet (low in protein), sheep fed a Lucerne-based diet (high in protein) harbored less nematodes at 56 days after infection. Supplementation of a hay-based diet with fish meal or soybean meal substantially reduced the worm burden in sheep infected with the nematodes Trichostrongylus colubriformis and Haemonchus contortus. It is postulated generally that protein supplementation enhances host immunity that prevents establishment or induces expulsion of parasites in both ruminants and non-ruminants.
Sponsor: Langston University, Langston, OK
PIs: Z. Wang, N. Wu, S. Hart, R. Merkel, L. J. Dawson, K. Matand, A. Goetsch, T. Sahlu
Insulin Resistance: Influence on Cytokine Regulation and Cell-mediated Immune Function
Insulin resistance occurs with many diseases including diabetes, obesity and aging thereby affecting a large proportion of the human population. Similar to humans, many horses are prone to obesity and insulin resistance. Insulin resistance has been associated with dysregulation of immune function. Infectious diseases occur more commonly in obese patients with insulin resistance. Immune failure and infection in the elderly may also be related to insulin resistance. We believe insulin resistance is associated with cytokine dysregulation and altered cell-mediated immunity in obese horses. We will compare immune function in blood cells from insulin resistant, obese horses and lean, healthy horses.
Sponsor: Research Advisory Committee, CVHS, OSU
PI: Todd C. Holbrook
Physiological Sciences: Dianne McFarlane
OKLAHOMA ANIMAL DISEASE DIAGNOSTIC LABORATORY
The Oklahoma Animal Disease Diagnostic Laboratory
The Oklahoma Animal Disease Diagnostic Laboratory provides accessible and accountable diagnostic service for Oklahoma veterinarians and animal owners in all 77 counties. Early detection of diseases provides the starting point for reducing their incidence and threat. The Laboratory also acts as a frontline sentinel for new and emerging diseases. OADDL promotes and protects the health and economic welfare of Oklahomans, supports the teaching and research missions of the OSU College of Veterinary Medicine, and conducts self-supported research aimed at developing more precise test procedures for commonly encountered, as well as emerging and foreign animal diseases, that may produce catastrophic losses (e.g., bovine viral diarrhea, malignant catarrhal fever, parvovirus disease, avian influenza, equine viral arteritis and encephalitis, and toxicoses related to oilfield wastes and agricultural chemicals). The Laboratory also conducts research and diagnostic tests for morbillivirus infections of marine mammals. The Laboratory, in conjunction with the Oklahoma State Department of Health, is involved with surveillance of West Nile fever in Oklahoma horses and birds. The Laboratory maintains full accreditation by the American Association of Veterinary Laboratory Diagnosticians.
PIs: Bill J. Johnson and staff