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Defective Interfering Rnas: Foes Of Viruses And Friends Of Virologists, Kunj Bihari Pathak, Peter D. Nagy

Plant Pathology Faculty Publications

Defective interfering (DI) RNAs are subviral RNAs produced during multiplication of RNA viruses by the error-prone viral replicase. DI-RNAs are parasitic RNAs that are derived from and associated with the parent virus, taking advantage of viral-coded protein factors for their multiplication. Recent advances in the field of DI RNA biology has led to a greater understanding about generation and evolution of DI-RNAs as well as the mechanism of symptom attenuation. Moreover, DI-RNAs are versatile tools in the hands of virologists and are used as less complex surrogate templates to understand the biology of their helper viruses. The ease of their …

Historical View And Physiology Demonstration At The Nmj Of The Crayfish Opener Muscle, Ann S. Cooper, Robin L. Cooper

Biology Faculty Publications

Here we present some of the key important discoveries made with the opener neuromuscular (NMJ) preparation of crustaceans and illustrate that there is still much to learn from this model preparation. In understanding the history one can appreciate why even today this NMJ still offers a rich playground to address questions regarding pre- and post-synaptic function and plasticity. The viability and ease of access to the terminal for intracellular as well as extracellular electrophysiology and imaging are significant advantages. The mechanisms behind the modulation of vesicular kinetics and fusion within the high- and low-output terminals are begging for investigation. The …

Expression Of Sorl1 And A Novel Sorl1 Splice Variant In Normal And Alzheimers Disease Brain, Karrie E. Grear, I-Fang Ling, James F. Simpson, Jennifer L. Furman, Christopher R. Simmons, Shawn L. Peterson, Frederick A. Schmitt, William R. Markesbery, Qiang Liu, Julia E. Crook, Steven G. Younkin, Guojun Bu, Steven Estus

Physiology Faculty Publications

BACKGROUND: Variations in sortilin-related receptor (SORL1) expression and function have been implicated in Alzheimers Disease (AD). Here, to gain insights into SORL1, we evaluated SORL1 expression and splicing as a function of AD and AD neuropathology, neural gene expression and a candidate single nucleotide polymorphism (SNP).

RESULTS: To identify SORL1 splice variants, we scanned each of the 46 internal SORL1 exons in human brain RNA samples and readily found SORL1 isoforms that lack exon 2 or exon 19. Quantification in a case-control series of the more abundant isoform lacking exon 2 (delta-2-SORL1), as well as the "full-length" SORL1 (FL-SORL1) isoform …

Aligned Nanotubule Membranes, Bruce J. Hinds Iii

Chemical and Materials Engineering Faculty Patents

A method is provided for producing a permeable membrane, comprising the steps of aligning a plurality of hollow nanotubules to form a mat, coating the mat with a continuous polymer matrix to form a membrane. The membrane is etched (a) to open the plurality of hollow nanotubules and form pores and (b) to oxidize the carboxyl groups to carboxylate groups. At least one additional functional unit having at least one available amine group to bind the at least one additional functional unit to the nanotubule end carboxylate group may be provided. Membranes fabricated in accordance with the method of the …

Aging Influences The Neural Correlates Of Lexical Decision But Not Automatic Semantic Priming, Brian T. Gold, Anders H. Andersen, Greg A. Jicha, Charles D. Smith

Neuroscience Faculty Publications

Human behavioral data indicate that older adults are slower to perform lexical decisions (LDs) than young adults but show similar reaction time gains when these decisions are primed semantically. The present study explored the functional neuroanatomic bases of these frequently observed behavioral findings. Young and older groups completed unprimed and primed LD tasks while functional magnetic resonance imaging (fMRI) was recorded, using a fully randomized trial design paralleling those used in behavioral research. Results from the unprimed task found that age-related slowing of LD was associated with decreased activation in perceptual extrastriate regions and increased activation in regions associated with …

Genetic Insulator For Preventing Influence By Another Gene Promoter, Susheng Gan, Mingtang Xie

Plant and Soil Sciences Faculty Patents

A 16 bp polynucleotide sequence of Arabidopsis thaliana is a genetic insulator that can effectively isolate a transgene from positional effects of neighboring gene activities in transgenic plant cells.

Methods And Devices For Cryopreservation Of Biological Cells And Tissues, Dayong Gao, Gary Van Zant, Xiangdong Cui

Mechanical Engineering Faculty Patents

Novel methods, compositions, and devices for achieving optimal cooling of living cells during cryopreservation are disclosed. In one aspect, the method comprises gradually cooling the cell to a first predetermined temperature, followed by rapidly cooling the cell to a second predetermined temperature. In another aspect, a device is described for achieving a desired cooling rate for a cell, comprising a first container for holding a cell, a second container for holding the first container, and optionally a frame for holding the first container in a spaced apart relationship with the second container. The method of the invention comprises placing cells …

An Eiav Field Isolate Reveals Much Higher Levels Of Subtype Variability Than Currently Reported For The Equine Lentivirus Family, Jodi K. Craigo, Shannon Barnes, Baoshan Zhang, Sheila J. Cook, Laryssa Howe, Charles J. Issel, Ronald C. Montelaro

Veterinary Science Faculty Publications

BACKGROUND: Equine infectious anemia virus (EIAV), a lentivirus that infects horses, has been utilized as an animal model for the study of HIV. Furthermore, the disease associated with the equine lentivirus poses a significant challenge to veterinary medicine around the world. As with all lentiviruses, EIAV has been shown to have a high propensity for genomic sequence and antigenic variation, especially in its envelope (Env) proteins. Recent studies have demonstrated Env variation to be a major determinant of vaccine efficacy, emphasizing the importance of defining natural variation among field isolates of EIAV. To date, however, published EIAV sequences have been …

The Leishmania Donovani Lipophosphoglycan Excludes The Vesicular Proton-Atpase From Phagosomes By Impairing The Recruitment Of Synaptotagmin V, Adrien F. Vinet, Mitsunori Fukuda, Salvatore J. Turco, Albert Descoteaux

Molecular and Cellular Biochemistry Faculty Publications

We recently showed that the exocytosis regulator Synaptotagmin (Syt) V is recruited to the nascent phagosome and remains associated throughout the maturation process. In this study, we investigated the possibility that Syt V plays a role in regulating interactions between the phagosome and the endocytic organelles. Silencing of Syt V by RNA interference revealed that Syt V contributes to phagolysosome biogenesis by regulating the acquisition of cathepsin D and the vesicular proton-ATPase. In contrast, recruitment of cathepsin B, the early endosomal marker EEA1 and the lysosomal marker LAMP1 to phagosomes was normal in the absence of Syt V. As Leishmania …

Measures Of Heart And Ventilatory Rates In Freely Moving Crayfish, Sonya M. Bierbower, Robin L. Cooper

Biology Faculty Publications

The fear, flight or fight response serves as the fundamental physiological basis for examining an organism's awareness of its environment under an impending predator attack. Although it is not known whether invertebrates posses an autonomic nervous system identical to that of vertebrates, evidence shows invertebrates have a sympathetic-like response to regulate the internal environment and ready the organism to act behaviorally to a given stimuli. Furthermore, this physiological response can be feasibly measured and it acts as a biological index for the animal's internal state. Measurements of the physiological response can be directly related to internal and external stressors through …

Inhibition Of Rna Recruitment And Replication Of An Rna Virus By Acridine Derivatives With Known Anti-Prion Activities, Zsuzsanna Sasvari, Stéphane Bach, Marc Blondel, Peter D. Nagy

Plant Pathology Faculty Publications

BACKGROUND: Small molecule inhibitors of RNA virus replication are potent antiviral drugs and useful to dissect selected steps in the replication process. To identify antiviral compounds against Tomato bushy stunt virus (TBSV), a model positive stranded RNA virus, we tested acridine derivatives, such as chlorpromazine (CPZ) and quinacrine (QC), which are active against prion-based diseases.

METHODOLOGY/PRINCIPAL FINDINGS: Here, we report that CPZ and QC compounds inhibited TBSV RNA accumulation in plants and in protoplasts. In vitro assays revealed that the inhibitory effects of these compounds were manifested at different steps of TBSV replication. QC was shown to have an effect …

Forage News [2009-10], Department Of Plant And Soil Sciences, University Of Kentucky

Forage News

• 10th Kentucky Grazing Conference
• 30th "Anniversary" Kentucky Alfalfa Conference
• KFGC Update
• Growing Alfalfa in the South
• Effects of a Freeze on Forages
• Grazing and Eliminative Behavior of Horses Grazing Bermudagrass
• Spatial Distribution of Livestock Concentration Areas and Soil Nutrients in Pastures
• Effect if Humate Application on the Forage Yield and Quality of Bermudagrass and Tall Fescue
• Effect of Repeated or One-Time Grazing of Kudzu by Sheep and Goats in Central Georgia
• Upcoming Events

Violence And Women's Mental Health: The Pain Unequalled, Part Two, Carol E. Jordan

Office for Policy Studies on Violence Against Women Publications

No abstract provided.

Transylvania’S Skull With A Hole In It, Charles T. Ambrose

Microbiology, Immunology, and Molecular Genetics Faculty Publications

No abstract provided.

Methods For Cancer Detection, Kimberly W. Anderson, Kimberly M.L. May, Leonidas G. Bachas

Chemical and Materials Engineering Faculty Patents

A method is provided for detecting an analyte indicative of a cancer or a metastatic disease condition, which utilizes the ability of the analyte to induce permeability in a barrier. The method includes providing a biosensor having a barrier which is substantially impermeable to an ion, a permeable membrane which is selective for the ion, and a detector capable of detecting the ion. The biosensor is contacted with a sample including at least one of the ion and the analyte, wherein the analyte causes at least a portion of the ion to pass through the barrier and the membrane. Passage …

Compositions Comprising Human Immunodeficiency Virus Tat Adsorbed To The Surface Of Anionic Nanoparticles, Russell J. Mumper, Jerold Woodward, Zhengrong Cui, Avindra Nath

Pharmaceutical Sciences Faculty Patents

Non-denatured, recombinant human immunodeficiency virus (HIV) Tat that is free of bacterial RNA and endotoxin is employed in an anti-HIV vaccine. A process of producing the recombinant Tat protein includes steps for removing bacterial RNA from the recombinant Tat and for removing endotoxin from the recombinant Tat protein. A Tat-adsorbed nanoparticle formulation and method of making the same. A method of vaccinating against and/or treating HIV infection comprises administering to a subject in need of such vaccination or treatment an immune-response inducing effective amount of the recombinant Tat protein.

Transcriptional Profiling Differences For Articular Cartilage And Repair Tissue In Equine Joint Surface Lesions, Michael J. Mienaltowski, Liping Huang, David D. Frisbie, C. Wayne Mcilwraith, Arnold J. Stromberg, Arne C. Bathke, James N. Macleod

Veterinary Science Faculty Publications

BACKGROUND: Full-thickness articular cartilage lesions that reach to the subchondral bone yet are restricted to the chondral compartment usually fill with a fibrocartilage-like repair tissue which is structurally and biomechanically compromised relative to normal articular cartilage. The objective of this study was to evaluate transcriptional differences between chondrocytes of normal articular cartilage and repair tissue cells four months post-microfracture.

METHODS: Bilateral one-cm2 full-thickness defects were made in the articular surface of both distal femurs of four adult horses followed by subchondral microfracture. Four months postoperatively, repair tissue from the lesion site and grossly normal articular cartilage from within the same …

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 …

Effects Of Genetic Deficiency Of Cyclooxygenase-1 Or Cyclooxygenase-2 On Functional And Histological Outcomes Following Traumatic Brain Injury In Mice, Matthew L. Kelso, Stephen W. Scheff, James R. Pauly, Charles D. Loftin

Pharmaceutical Sciences Faculty Publications

BACKGROUND: Neuroinflammation contributes to the pathophysiology of acute CNS injury, including traumatic brain injury (TBI). Although prostaglandin lipid mediators of inflammation contribute to a variety of inflammatory responses, their importance in neuroinflammation is not clear. There are conflicting reports as to the efficacy of inhibiting the enzymes required for prostaglandin formation, cyclooxygenase (COX) -1 and COX-2, for improving outcomes following TBI. The purpose of the current study was to determine the role of the COX isoforms in contributing to pathological processes resulting from TBI by utilizing mice deficient in COX-1 or COX-2.

RESULTS: Following a mild controlled cortical impact injury, …

The Genome Of Nectria Haematococca: Contribution Of Supernumerary Chromosomes To Gene Expansion, Jeffrey J. Coleman, Steve D. Rounsley, Marianela Rodriguez-Carres, Alan Kuo, Catherine C. Wasmann, Jane Grimwood, Jeremy Schmutz, Masatoki Taga, Gerard J. White, Shiguo Zhou, David C. Schwartz, Michael Freitag, Li-Jun Ma, Etienne G. J. Danchin, Bernard Henrissat, Pedro M. Coutinho, David R. Nelson, Dave Straney, Carolyn A. Napoli, Bridget M. Barker, Michael Gribskov, Martijn Rep, Scott Kroken, István Molnár, Christopher Rensing, John C. Kennell, Jorge Zamora, Mark L. Farman, Eric U. Selker, Asaf Salamov, Harris Shapiro, Jasmyn Pangilinan, Erika Lindquist, Casey Lamers, Igor V. Grigoriev, David M. Geiser, Sarah F. Covert, Esteban Temporini, Hans D. Vanetten

Plant Pathology Faculty Publications

The ascomycetous fungus Nectria haematococca, (asexual name Fusarium solani), is a member of a group of >50 species known as the "Fusarium solani species complex". Members of this complex have diverse biological properties including the ability to cause disease on >100 genera of plants and opportunistic infections in humans. The current research analyzed the most extensively studied member of this complex, N. haematococca mating population VI (MPVI). Several genes controlling the ability of individual isolates of this species to colonize specific habitats are located on supernumerary chromosomes. Optical mapping revealed that the sequenced isolate has 17 chromosomes ranging from 530 …

Tumor Necrosis Factor-Alpha (Tnf-Α) Enhances Functional Thermal And Chemical Responses Of Trp Cation Channels In Human Synoviocytes, Mikhail Y. Kochukov, Terry A. Mcnearney, Huaizhi Yin, Liping Zhang, Fei Ma, Larissa Ponomareva, Sarah Abshire, Karin N. Westlund

Physiology Faculty Publications

Background: We have shown functional expression of several TRP channels on human synovial cells, proposing significance in known calcium dependent proliferative and secretory responses in joint inflammation. The present study further characterizes synoviocyte TRP expression and activation responses to thermal and osmotic stimuli after pre-treatment with proinflammatory mediator tumor necrosis factor alpha (TNF-α, EC50 1.3221 x 10^-10 g/L).

Results: Fluorescent imaging of Fura-2 loaded human SW982 synoviocytes reveals immediate and delayed cytosolic calcium oscillations elicited by (1) TRPV1 agonists capsaicin and resiniferatoxin (20-40% of cells), (2) moderate and noxious temperature change, and (3) osmotic stress TRPV4 activation (11.5% of …

Integration Of Irradiation With Cytoplasmic Incompatibility To Facilitate A Lymphatic Filariasis Vector Elimination Approach, Corey L. Brelsfoard, William St. Clair, Stephen L. Dobson

Entomology Faculty Publications

BACKGROUND: Mass drug administration (MDA) is the emphasis of an ongoing global lymphatic filariasis (LF) elimination program by the World Health Organization, in which the entire 'at risk' human population is treated annually with anti-filarial drugs. However, there is evidence that the MDA strategy may not be equally appropriate in all areas of LF transmission, leading to calls for the augmentation of MDA with anti-vector interventions. One potential augmentative intervention is the elimination of vectors via repeated inundative releases of male mosquitoes made cytoplasmically incompatible via an infection with Wolbachia bacteria. However, with a reduction in the vector population size, …

Muc1 Is A Downstream Target Of Stat3 And Regulates Lung Cancer Cell Survival And Invasion, Jingchun Gao, Matthew J. Mcconnell, Bin Yu, Jiannong Li, Justin M. Balko, Esther P. Black, Joseph O. Johnson, Mark C. Lloyd, Soner Altiok, Eric B. Haura

Pharmaceutical Sciences Faculty Publications

Signal transducer and activator of transcription 3 (STAT3) is aberrantly activated in human cancer including lung cancer and has been implicated in transformation, tumorigenicity, and metastasis. One putative downstream gene regulated by Stat3 is MUC1 which also has important roles in tumorigenesis. We determined if Stat3 regulates MUC1 in lung cancer cell lines and what function MUC1 plays in lung cancer cell biology. We examined MUC1 expression in non-small cell lung cancer (NSCLC) cell lines and found high levels of MUC1 protein expression associated with higher levels of tyrosine phosphorylated STAT3. STAT3 knockdown downregulated MUC1 expression whereas constitutive STAT3 expression …

Suppression Of Rft1 Expression Does Not Impair The Transbilayer Movement Of Man5glcnac2-P-P-Dolichol In Sealed Microsomes From Yeast., Jeffrey S. Rush, Ningguo Gao, Mark A Lehrman, Sergey Matveev, Charles J Waechter

Molecular and Cellular Biochemistry Faculty Publications

To further evaluate the role of Rft1 in the transbilayer movement of Man(5)GlcNAc(2)-P-P-dolichol (M5-DLO), a series of experiments was conducted with intact cells and sealed microsomal vesicles. First, an unexpectedly large accumulation (37-fold) of M5-DLO was observed in Rft1-depleted cells (YG1137) relative to Glc(3)Man(9)GlcNAc(2)-P-P-Dol in wild type (SS328) cells when glycolipid levels were compared by fluorophore-assisted carbohydrate electrophoresis analysis. When sealed microsomes from wild type cells and cells depleted of Rft1 were incubated with GDP-[(3)H]mannose or UDP-[(3)H]GlcNAc in the presence of unlabeled GDP-Man, no difference was observed in the rate of synthesis of [(3)H]Man(9)GlcNAc(2)-P-P-dolichol or Man(9)[(3)H]GlcNAc(2)-P-P-dolichol, respectively. In addition, no …

Rapid Fixation Of Non-Native Alleles Revealed By Genome-Wide Snp Analysis Of Hybrid Tiger Salamanders, Benjamin M. Fitzpatrick, Jarrett R. Johnson, D. Kevin Kump, H. Bradley Shaffer, Jeramiah J. Smith, S. Randal Voss

Biology Faculty Publications

BACKGROUND: Hybrid zones represent valuable opportunities to observe evolution in systems that are unusually dynamic and where the potential for the origin of novelty and rapid adaptation co-occur with the potential for dysfunction. Recently initiated hybrid zones are particularly exciting evolutionary experiments because ongoing natural selection on novel genetic combinations can be studied in ecological time. Moreover, when hybrid zones involve native and introduced species, complex genetic patterns present important challenges for conservation policy. To assess variation of admixture dynamics, we scored a large panel of markers in five wild hybrid populations formed when Barred Tiger Salamanders were introduced into …

Evolution Of A Subtilisin-Like Protease Gene Family In The Grass Endophytic Fungus Epichloë Festucae, Michelle K. Bryant, Christopher L. Schardl, Uljana Hesse, Barry Scott

Plant Pathology Faculty Publications

BACKGROUND: Subtilisin-like proteases (SLPs) form a superfamily of enzymes that act to degrade protein substrates. In fungi, SLPs can play either a general nutritive role, or may play specific roles in cell metabolism, or as pathogenicity or virulence factors.

RESULTS: Fifteen different genes encoding SLPs were identified in the genome of the grass endophytic fungus Epichloë festucae. Phylogenetic analysis indicated that these SLPs belong to four different subtilisin families: proteinase K, kexin, pyrolysin and subtilisin. The pattern of intron loss and gain is consistent with this phylogeny. E. festucae is exceptional in that it contains two kexin-like genes. Phylogenetic analysis …

Lipid Nanoparticles With Accessible Nickel As A Vaccine Delivery System For Single And Multiple His-Tagged Hiv Antigens, Weili Yan, Anekant Jain, Ronan O'Carra, Jerold Woodward, Wenxue Li, Guanhan Li, Avindra Nath, Russell J. Mumper

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Lipid-based nanoparticles (NPs) with a small amount of surface-chelated nickel (Ni-NPs) were developed to easily formulate the HIV his-tagged Tat protein, as well as to formulate and co-deliver two HIV antigens (his-p24 and his-Nef) on one particle. Female BALB/c mice were immunized by s.c. injection with his-Tat/Ni-NP formulation (1.5 μg Tat-his/mouse) and control formulations on day 0 and 14. The day 28 anti-Tat specific IgG titer with his-Tat/Ni-NP was significantly greater than that with Alum/his-Tat. Furthermore, splenocytes from his-Tat/Ni-NP immunized mice secreted significantly higher IFN-γ than those from mice immunized with Alum/his-Tat. Although Ni-NPs did not show better adjuvant activity …

Predictive Archaeological Modeling Using Gis-Based Fuzzy Set Estimation: A Case Study In Woodford County, Kentucky, Philip B. Mink, John Ripy, Keiron Bailey, Ted H. Grossardt

Kentucky Transportation Center Faculty and Researcher Publications

Analytic predictive archaeological models can have great utility for state Departments of Transportation, but it is difficult to model the likelihood of prehistoric settlement using geographical proxy predictor variables because of the complexity of how settlement choices were actually made, and the complex interaction between these variables using GIS. In many cases classic statistical modeling approaches require too much data to be useful. This research reports on a preliminary predictive model that combines Spatial Analyst and fuzzy logic modeling to capture expert archaeological knowledge and convert this into predictive surface. A test area was defined in Woodford County, KY and …

Dna-Binding By Haemophilus Influenzae And Escherichia Coli Ybab, Members Of A Widely-Distributed Bacterial Protein Family, Anne E. Cooley, Sean P. Riley, Keith Kral, M. Clarke Miller, Edward Demoll, Michael G. Fried, Brian Stevenson

Microbiology, Immunology, and Molecular Genetics Faculty Publications

BACKGROUND: Genes orthologous to the ybaB loci of Escherichia coli and Haemophilus influenzae are widely distributed among eubacteria. Several years ago, the three-dimensional structures of the YbaB orthologs of both E. coli and H. influenzae were determined, revealing a novel "tweezer"-like structure. However, a function for YbaB had remained elusive, with an early study of the H. influenzae ortholog failing to detect DNA-binding activity. Our group recently determined that the Borrelia burgdorferi YbaB ortholog, EbfC, is a DNA-binding protein. To reconcile those results, we assessed the abilities of both the H. influenzae and E. coli YbaB proteins to bind DNA …

Distinctive Interactions Of The Arabidopsis Homolog Of The 30 Kd Subunit Of The Cleavage And Polyadenylation Specificity Factor (Atcpsf30) With Other Polyadenylation Factor Subunits, Suryadevara Rao, Randy D. Dinkins, Arthur G. Hunt

Plant and Soil Sciences Faculty Publications

BACKGROUND: The Arabidopsis ortholog of the 30 kD subunit of the mammalian Cleavage and Polyadenylation Specificity Factor (AtCPSF30) is an RNA-binding endonuclease that is associated with other Arabidopsis CPSF subunits (orthologs of the 160, 100, and 73 kD subunits of CPSF). In order to further explore the functions of AtCPSF30, the subcellular distribution of the protein was examined by over-expressing fusion proteins containing fluorescent reporters linked to different CPSF subunits.

RESULTS: It was found that AtCPSF30 by itself localizes, not to the nucleus, but to the cytoplasm. AtCPSF30 could be found in the nucleus when co-expressed with AtCPSF160 or AtCPSF73(I), …