Life Sciences Commons^™

Gene Expression Regulators Lin-11 And Let-711, In Modulating The Rate Of Aging And Lifespan, In C. Elegans., Tseten Yeshi Jamling

University of Kentucky Doctoral Dissertations

lin-11 and let-711 are early-developmental gene expression regulators with no previously known roles in aging regulation. Yet, they show strong aging-correlated expression profiles (Lund, Tedesco et al. 2002). lin-11 is strongly upregulated in very old worm populations, and let-711 is progressively downregulated in aging worm populations. Microarray studies were performed to identify their genome-wide targets, which were then subjected to further lifespan and genetic analysis to investigate their role in C. elegans aging.

The results indicate that the target pools of both lin-11 and let-711 are enriched for aging genes, since a significant number of tested genes increased lifespan. This …

Biological Control Of The Black Cutworm, Agrotis Ipsilon (Lepidoptera: Noctuidae), And Endophyte Mediated Tritrophic Interactions In Turfgrass, Andrea Jeanne Bixby-Brosi

University of Kentucky Doctoral Dissertations

Components of successful pest management programs must be complementary and not antagonistic. This project examined interactions between natural enemies of the black cutworm, Agrotis ipsilon (Hufnagel), an important turfgrass pest, and host plant resistance by endophytic grass.

Agrotis ipsilon nucleopolyhedrovirus (AgipMNPV) was examined as a bio-insecticide for controlling A. ipsilon in turfgrass. Fresh (1-week-old) AgipMNPV residues killed 76−86% of neonates hatching from eggs on golf course tees, however, residual control of implanted larvae lasted no more than a few weeks. Combinations of AgipMNPV with adjuvants, such as optical brightener and lignin, failed to accelerate or extend …

Engineering Proteins With Unique Characteristics For Diagnostics And Biosensors, Smita Joel

University of Kentucky Doctoral Dissertations

Proteins possess a broad range of structural and functional properties and, therefore, can be employed in a variety of biomedical applications. While a good number of protein-based biosensing systems and biosensors for target analytes have been developed, the search for versatile, highly sensitive and selective sensors with long term stability able to provide fast detection of target analytes continues to be a challenge. To that end, we now report the design and development of modified proteins with tailored characteristics and their further utilization in the development of biosensing systems.

We take advantage of binding proteins that undergo a change in …

Loss Of Bloom Syndrome Protein Causes Destabilization Of Genomic Architecture And Is Complemented By Ectopic Expression Of Escherichia Coli Recg In Human Cells, Michael Wayne Killen

University of Kentucky Doctoral Dissertations

Genomic instability driven by non-allelic homologous recombination (NAHR) provides a realistic mechanism that could account for the numerous chromosomal abnormalities that are hallmarks of cancer. We recently demonstrated that this type of instability could be assayed by analyzing the copy number variation of the human ribosomal RNA gene clusters (rDNA). Further, we found that gene cluster instability (GCI) was present in greater than 50% of the human cancer samples that were tested. Here, data is presented that confirms this phenomenon in the human GAGE gene cluster of those cancer patients. This adds credence to the hypothesis that NAHR could be …

Using Linked Household-Level Datasets To Explain Consumer Response To Bse In Canada, Xin Wang

University of Kentucky Doctoral Dissertations

Household-level Canadian meat purchases from 2002-2008, a Food Opinions Survey conducted in 2008 at the national level and household-level egg purchases from 2002-2005 in Alberta and Ontario were used to explore consumer responses to Bovine Spongiform Encephalopathy (BSE) in Canada.

The opinions survey focused on nutritional priorities, general and specific food safety concerns, and trust in government and food industry decision makers. The egg data set contained specific product information allowing us to distinguish purchases of conventional eggs from those of value-added eggs with perceived health attributes. Thus, the egg purchase data appeared to be an interesting proxy of revealed …

Role Of Pi3k-Akt Pathway In The Age Associated Decline In Tlr Mediated Activation Of Innate And Adaptive Immune Responses, Mosoka Papa Fallah

University of Kentucky Doctoral Dissertations

Immunosenescence results in reduced immune response to infections with Streptococcus pneumoniae as well as to pneumococcal polysaccharide vaccines. The antibody response to the capsular polysaccharide (CPS) provides protection against S. pneumoniae infection. CPS immunoresponse is T cell independent and needs the macrophage-derived cytokines such as IL-12, IL-6 and IL-1β to elicit an antibody response. We showed a cytokine dysregulation, i.e. a decrease in IL-12, IL-6 and TNF-α but an increase in IL-10, in the aged (18-24 months old comparable to >65 years in human) compared to young adult mouse (8-12 weeks less than 65 years old) splenic macrophages (SM) or …

Post-Transcriptional Regulation Of Afp And Igm Genes, Lilia M. Turcios

University of Kentucky Doctoral Dissertations

Gene expression can be regulated at multiple steps once transcription is initiated. I have studied two different gene models, the α-Fetoprotein (AFP) and the immunoglobulin heavy chain (IgM) genes, to better understand post-transcriptional gene regulation mechanisms. The AFP gene is highly expressed during fetal liver development and dramatically repressed after birth. There is a mouse strain-specific difference between adult levels of AFP, with BALB/cJ mice expressing 10 to 20-fold higher levels compared to other mouse strains. BALB/cJ mice express low levels of Zhx2 and thus incompletely repress AFP. Despite differences in steady state AFP mRNA levels in the adult liver …

Using Manual Defoliation To Simulate Soybean Rust: Effect On Growth And Yield Formation, Abdullah Mohammad Aqeel

University of Kentucky Doctoral Dissertations

Field experiments were conducted in Kentucky and Louisiana in 2008 and 2009 (split-plot in a randomized complete block design with four replications) to investigate it is possible to simulate with manual defoliation the effect of soybean rust (SBR) (Phakopsora pachyrhizi Syd. and P. Syd) injury on a healthy soybean [Glycine max, (L.) Merr.] canopy, understand how defoliation affects the growth dynamics and canopy light interception, and if defoliation affectsleaf senescence and nitrogen remobilization during the seed-filling period. Two manual defoliation treatments based on changes in effective leaf area index (ELAI) (calculated as the reduction in leaf area …

Efficacy Of Organically Certifiable Materials And Natural Compounds Against Foliar Hemibiotrophic And Necrotrophic Fungi In Cantaloupe And Tomato, Merari Feliciano-Rivera

University of Kentucky Doctoral Dissertations

Kentucky reported a solid 13.1% growth in certified organic land from 1997 to 2002. The relative lack of research on disease management practices in Kentucky consistent with organic regulations is an issue that needs to be addressed to provide more reliable information to local farmers. Thus, the first objective of this research was to investigate the potential disease control obtained with natural, organically certifiable spray materials against Colletotrichum orbiculare in vitro and in vivo. The second objective was to test certifiable spray materials in combinations to identify synergistic interactions. The third objective was to evaluate Organic Material Review Institute …

Human Chondrosarcoma Cells Acquire An Epithelial-Like Gene Expression Pattern Via An Epigenetic Switch: Evidence For Mesenchymal-Epithelial Transition During Sarcomagenesis., Matthew P. Fitzgerald, Francoise Gourronc, Melissa L.T. Teoh, Matthew J. Provenzano, Adam J. Case, James A. Martin, Frederick E. Domann

Journal Articles: Cellular & Integrative Physiology

Chondrocytes are mesenchymally derived cells that reportedly acquire some epithelial characteristics; however, whether this is a progression through a mesenchymal to epithelial transition (MET) during chondrosarcoma development is still a matter of investigation. We observed that chondrosarcoma cells acquired the expression of four epithelial markers, E-cadherin,desmocollin 3, maspin, and 14-3-3σ, all of which are governed epigenetically through cytosine methylation. Indeed, loss of cytosine methylation was tightly associated with acquired expression of both maspin and 14-3-3σ in chondrosarcomas. In contrast, chondrocyte cells were negative for maspin and 14-3-3σ and displayed nearly complete DNA methylation. Robust activation of these genes was also …

Genotoxin-Induced Acetylation Of The Werner Syndrome Protein (Wrn) And Effect On Its Dna Metabolic Function, Enerlyn Meliza Lozada Santiago

University of Kentucky Doctoral Dissertations

Loss of function of the WRN protein causes the genetic disorder Werner Syndrome that is characterized by increased cancer and premature aging. WRN belongs to the RecQ helicase family that plays key roles in preventing genome instability. In response to treatment with genotoxins, WRN is subject to post-translational modification. The relationship of post-translational modification of WRN with its function in DNA metabolism is unknown. There is accumulating evidence suggesting that WRN contributes to the maintenance of genomic integrity through its involvement in DNA replication. Consistent with this notion, WS cells are sensitive to DNA replication inhibitors and DNA damaging agents …

Modeling And Quantitative Analysis Of White Matter Fiber Tracts In Diffusion Tensor Imaging, Xuwei Liang

University of Kentucky Doctoral Dissertations

Diffusion tensor imaging (DTI) is a structural magnetic resonance imaging (MRI) technique to record incoherent motion of water molecules and has been used to detect micro structural white matter alterations in clinical studies to explore certain brain disorders. A variety of DTI based techniques for detecting brain disorders and facilitating clinical group analysis have been developed in the past few years. However, there are two crucial issues that have great impacts on the performance of those algorithms. One is that brain neural pathways appear in complicated 3D structures which are inappropriate and inaccurate to be approximated by simple 2D structures, …

Identification Of Activities Involved In Cag/Ctg Repeat Instability, Nelson Lap Shun Chan

University of Kentucky Doctoral Dissertations

CAG/CTG repeat instability is associated with at least 14 neurological disorders, including Huntington’s disease and Myotonic dystrophy type 1. In vitro and in vivo studies have showed that CAG/CTG repeats form a stable hairpin that is believed to be the intermediate for repeat expansion and contraction.

Addition of extra DNA is essential for repeat expansion, so DNA synthesis is one of the keys for repeat expansion. In vivo studies reveal that 3’ CTG slippage with subsequent hairpin formation (henceforth called the 3’ CTG slippage hairpin) occurs during DNA synthesis. It is proposed that hairpin tolerance machinery is activated because prolonged …

Fabrication And Characterization Of Mesoscale Protein Patterns Using Atomic Force Microscopy (Afm), Pei Gao

University of Kentucky Doctoral Dissertations

A versatile AFM local oxidation lithography was developed for fabricating clean protein patterns ranging from nanometer to sub-millimeter scale on octadecyltrichlorosilane (OTS) layer of Si (100) wafer. This protein patterning method can generate bio-active protein pattern with a clean background without the need of the anti-fouling the surface or repetitive rinsing.

As a model system, lysozyme protein patterns were investigated through their binding reactions with antibodies and aptamers by AFM. Polyclonal anti-lysozyme antibodies and anti-lysozyme aptamer are found to preferentially bind to the lysozyme molecules on the edge of a protein pattern before their binding to the interior ones. It …

Role Of Lipids In Tombusvirus Replication, Monika Sharma

University of Kentucky Doctoral Dissertations

Positive-strand RNA virus group are the most abundant among viruses affecting plants and animals. To successfully achieve replication, these viruses usurp or co-opt host proteins. To facilitate the discovery of host factors involved in Tomato bushy stunt virus (TBSV), yeast has been developed as a surrogate model host. Genome-wide approaches covering 95% of yeast genes, has revealed approximately hundred factors that could affect virus replication. Among the identified host factors, there are fourteen yeast genes, which affect/regulate lipid metabolism of the host.

One of the identified host gene is ERG25, which is an important factor for sterol biosynthesis pathway, affecting …

Chronic Pulmonary Cavitary Tuberculosis In Rabbits: A Failed Host Immune Response, Selvakumar Subbian, Liana Tsenova, Guibin Yang, Paul O'Brien, Sven Parsons, Blas Peixoto, Leslie Taylor, Dorothy Fallows, Gilla Kaplan

Publications and Research

The molecular determinants of the immune response to Mycobacterium tuberculosis HN878 infection in a rabbit model of pulmonary cavitary tuberculosis were studied. Aerosol infection of rabbits resulted in a highly differentially expressed global transcriptome in the lungs at 2 weeks, which dropped at 4 weeks and then gradually increased. While IFNγ was progressively upregulated throughout the infection, several other genes in the IFNγ network were not. T-cell activation network genes were gradually upregulated and maximally induced at 12 weeks. Similarly, the IL4 and B-cell activation networks were progressively upregulated, many reaching high levels between 12 and 16 weeks. Delayed peak …

A Messenger Molecule Governs Interdependency In An Evolved Pseudomonas Aeruginosa Biofilm Community, Thomas Macelliott Johnson

Honors Theses and Capstones

Biofilm populations are known to harbor great diversity, but the importance of this diversity is not fully understood. A likely contributor to this variation is the second messenger molecule cyclic-di-GMP: low levels associate with a planktonic lifestyle while high levels favor biofilm formation. In ongoing studies of an evolving biofilm population of Pseudomonas aeruginosa (PA), we observed extensive ecological diversification that may relate to this lifestyle switch. In PA, the gene bifA encodes a phosphodiesterase that is known to degrade cyclic-di-GMP and reduce biofilm. This gene was cloned onto a plasmid under control of an inducible promoter and the plasmid …

Total Mercury In Stranded Marine Mammals From The Oregon And Southern Washington Coasts, Nathan J. Wintle

Dissertations and Theses

Muscle samples from 105 marine mammals stranded along the Oregon-Washington coasts (2002-2009) were tested for levels of total mercury by Cold Vapor Atomic Fluorescence Spectrometry. The total mercury present is predominantly in the form of highly toxic methylmercury. After normalizing muscle tissue to 75% water weight, due to variance in water content, Steller sea lions (Eumetopias jubatus) and northern elephant seals (Mirounga angustirostris) exhibited the highest mean concentrations of total mercury followed by harbor seals (Phoca vitulina), harbor porpoises (Phocoena phocoena), and California sea lions (Zalophus californianus); 0.34 ± 0.278, …

Molecular Genetic Analysis Of Creld1 In Patients With Heterotaxy Disorder, Samaneh Zhian

Dissertations and Theses

Heterotaxy refers to the abnormal arrangement of internal organs in relation to each other. Model organism studies have shown that functions of more than eighty genes are required for normal asymmetric left-right organ development. CRELD1 has been shown to be necessary for proper heart development and mutations in CRELD1 are known to increase risk of cardiac atrioventricular septal defects (AVSD). AVSD is the most common form of heart defect associated with heterotaxy, and we have previously shown that some individuals heterotaxy-related AVSD have mutations in CRELD1.

Therefore, we propose to examine the CRELD1 gene in a large sample of …

Using Archaeological Fish Remains To Determine The Native Status Of Anadromous Salmonids In The Upper Klamath Basin (Oregon, Usa) Through Mtdna And Geochemical Analysis, Alexander E. Stevenson

Dissertations and Theses

Within the Upper Klamath Basin, Oregon, the native status of anadromous salmonids (Oncorhynchus spp.) has been a long standing question. Ongoing efforts to establish if these fish were native to the region prior to the construction of the Copco I Dam on the Klamath River (c.1917) have relied on sparse, contradictory and sometimes unreliable historic documentation and informant testimony. Current restoration projects with very high financial and social costs necessitate accurate and reliable data on salmonid species which once called the region home. Often, archaeolofaunal remains present a novel way to determine species present in an area prior to …

Characteristics Of Road-Kill Locations Of San Clemente Island Foxes, Nathan P. Snow, William F. Andelt, Nicholas P. Gould

United States Department of Agriculture Wildlife Services: Staff Publications

Mortalities from collisions with vehicles have created concern for the welfare of the San Clemente Island fox (Urocyon littoralis clementae); 1 of only 6 genetically distinct subspecies of island fox. To find possible solutions for minimizing these mortalities, we compared 9 characteristics of roads and roadsides at kill-sites and control-sites to ascertain whether certain features were associated with risk of collisions. We found that kill-sites were positively associated with the volume of traffic, and negatively associated with the distance of motorists’ visibility, which had not been previously identified for island foxes. Additionally, visual obstructions along roadsides (i.e., steep …

Postfledging Survival And Habitat Use Of Spotted Towhees (Pipilo Maculatus) In An Urban Park, Amy Ann Shipley

Dissertations and Theses

Habitat fragmentation, and the resulting increase in edge habitat, has important effects on birds, including the increased probability of nest predation, changes in habitat structure, and the increased presence of non-native plant species. It is critical to understand the effects of fragmentation at all stages of the avian life cycle, including the often overlooked postfledging period. Because much of juvenile mortality occurs during the immediate postfledging period, and juvenile mortality contributes substantially to population dynamics, it is necessary to understand if fledgling survival is reduced in edge habitats and if fledglings' survival is influenced by their habitat use. During 2008 …

Interception In Open-Grown Douglas-Fir (Pseudotsuga Menziesii) Urban Canopy, Mitchell Bixby

Dissertations and Theses

I hypothesized that Douglas-fir trees (Pseudotsuga menziesii) standing apart from other trees ('open-grown') will intercept more rainfall than Douglas-fir trees standing near other trees ('closed-canopy'). Open-grown trees differ structurally and are more common in urban settings, yet have been infrequently studied. Existing literature, based primarily on closed-canopy trees, suggests Douglas-fir trees in Pacific Northwest forests intercept approximately 25% of rainfall annually. Because open-grown trees have more vertical leaf area than individual trees in closed-canopy forests, I expected to find higher interception by open-grown trees.

I collected throughfall under four open-grown Douglas-firs using six static collectors ('buckets') per tree, …

Population Structure Of Island-Associated Pantropical Spotted Dolphins (Stenella Attenuata) In Hawaiian Waters, Sarah Shelby Courbis

Dissertations and Theses

Understanding gene flow, diversity, and dispersal patterns is important for predicting effects of natural events and anthropogenic activities on dolphin populations. With the very recent exceptions of false killer whales (Pseudorca crassidens), spinner dolphins (Stenella longirostris), and common bottlenose dolphins (Tursiops truncatus), Hawaiian odontocete species are managed as single stocks within the U.S. Hawaiian Exclusive Economic Zone. These exceptions are a result of recent studies that have indicated that some species have populations that show fidelity to individual islands or groups of islands, resulting in genetic differentiation, often with management implications. The first part …

Effects Of Lateral Ligament Sectioning On The Stability Of The Ankle And Subtalar Joint, Stacie I. Ringleb, Ajaya Dhakal, Claude D. Anderson, Sebastian Bawab, Rajesh Paranjape

Mechanical & Aerospace Engineering Faculty Publications

Patients with subtalar joint instability are often diagnosed with ankle instability. Only after a prolonged period of time in which a patient does not improve after treatment for ankle instability is subtalar joint instability considered. To develop a clinically relevant method to diagnose subtalar joint instability, the kinematics of the simulated unstable subtalar joint were examined. A 6 degree-of-freedom positioning and loading device was developed. Plantarflexion/dorsiflexion, inversion/eversion, and internal/external rotation were applied individually or as coupled motions along with an anterior/posterior drawer. Kinematic data were collected from sensors attached to the calcaneus, talus, and tibia by keeping all the ligaments …

Identification And Characterization Of Fkbp52-Specific Inhibitors For The Treatment Of Prostate Cancer, Johanny Tonos De Leon

Open Access Theses & Dissertations

Steroid hormone receptors require the ordered assembly of various chaperone and cochaperone proteins in order to reach a functional state. The final stage in the receptor maturation process requires the formation of a mutimeric complex consisting of Hsp90 dimer, p23, and one of several large immunophilins. Studies conducted previously demonstrated that the large immunophilin FKBP52 acts to potentiate glucocorticoid, androgen, and progesterone receptor signaling pathways. The aim of these studies was to identify and characterize FKBP52-specific inhibitors that would not only serve as tools for the pharmacological analysis of FKBP52-receptor interactions, but may also lead to novel drugs with significant …

Development Of A Novel Metallo-Lipid Microparticle Delivery System For A Leishmania Mexicana Dna Vaccine Candidate, Joanna B. Valencia

Open Access Theses & Dissertations

ABSTRACT

Background and Significance.

The leishmaniases are an important group of neglected tropical parasitic diseases caused by parasitic protozoa of the genus Leishmania. International health authorities estimate that 350 million people around the world are at risk for contracting leishmaniasis. Two million new cases occur each year and 12 million people are presently infected worldwide. Leishmaniasis is endemic throughout the Americas region including all of Central America, most of South America, Mexico and southern Texas. Leishmania is also considered a threat to the military readiness of U.S. troops deployed to the Middle East or Latin America.

First- and second-line treatments …

Causes And Consequences Of Changes In Algal Community Structure Of Arctic Tundra Ponds From Barrow, Alaska, Mariana Vargas Medrano

Open Access Theses & Dissertations

Algal communities in the Arctic are highly sensitive to environmental change, such as longer growing seasons and changes in ice cover. On the north slope of Alaska, ponds and lakes cover large portions of the total land area. We surveyed the periphytic algal communities in tundra ponds from Barrow, Alaska to assess environmental effects on their populations. Samples were collected in August 2008-10 from two main study areas: the IBP (the International Biological Program), which was also studied in 1970-71 but now is located near to human settlements, and the BEO (Barrow Environmental Observatory), which is within a protected area. …

Phylogeography Of Northern Populations Of The Black-Tailed Rattlesnake (Crotalus Molossus Baird And Girard, 1853) With The Revalidation Of C. Ornatus Hallowell, 1854, Christopher Anderson

Open Access Theses & Dissertations

The black-tailed rattlesnake (Crotalus molossus Baird and Girard, 1853) is a wide-ranging species complex with representatives from the southwestern United States through the highlands of central and southern Mexico. The systematics of this group has received little attention and in the past six decades only two taxonomic revisions have been proposed (the transfer of C. basiliscus oaxacus to C. molossus and the elevation of C. m. estebanensis to full species). However, a recent revision of the Neotropical rattlesnakes (C. durissus and C. simus) recovered a polyphyletic C. molossus. Sequenced data were obtained from three mitochondrial genes …

Prediction Of Ribonucleic Acid Secondary Structures Using A Heuristic Backtracking Search, Christopher Roman Cuellar

Open Access Theses & Dissertations

Ribonucleic acid (RNA) is essential for all forms of life. RNA is made up of a large chain of nucleotide bases: Guanine (G), Uracil (U), Cytosine (C), and Adenine (A). An RNA strand can fold on itself to allow G-C, A-U, and G-U bases to form hydrogen bonds, this is known as a secondary structure. Knowing the secondary structure of an RNA chain is very important because it will allow researchers to better understand its specific functions. RNA will create secondary structures that tend to minimize their free energy. RNA secondary structure prediction is the attempt to predict physical folding …