Physical Sciences and Mathematics Commons^™

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 …

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, …

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 …

The Ecology Of Disturbances And Global Change In The Montane Grasslands Of The Nilgiris, South India, Madhusudan P. Srinivasan

University of Kentucky Doctoral Dissertations

Biodiversity rich regions worldwide face threats from various global change agents. This research quantifies environmental influences on vegetation, and the impacts of exotic woody plant invasion and anthropogenic nitrogen (N) deposition in a global biodiversity hotspot. The study was conducted in the montane grasslands of the Nilgiris, Western Ghats, and outlines potential management options for this region. Specifically, I examined (1) the role of environmental factors in influencing native plant distribution and ecosystem properties, (2) the status and impact of exotic shrub (Scotch broom, henceforth broom) invasion, (3) the role of disturbances in the success of broom, (4) the role …

Analysis Of Differential Gene Expression And Alternative Splicing In The Liver And Gastrointestinal Tract In The Lactating Rat, Antony Thomas Athippozhy

University of Kentucky Doctoral Dissertations

Rat exon microarrays were utilized to detect changes in mRNA expression and alternative splicing in the liver, duodenum, jejunum, and ileum of the lactating rat when compared to age-matched virgin controls. Analysis of data at the level of gene expression revealed differential expression of genes involved in cholesterol biosynthesis in each tissue examined, suggesting increased Sterol Response Element Binding Protein activity. We also detected decreased mRNA from components of the T-cell signaling pathway in the jejunum and ileum. We characterized expression of solute carrier and adenosine triphosphate binding cassette proteins. In addition to characterizing genes by pathway, we have also …

Biochemical Characterization Of Human Mismatch Recognition Proteins Mutsα And Mutsβ, Lei Tian

University of Kentucky Doctoral Dissertations

The integrity of an organism's genome depends on the fidelity of DNA replication and the efficiency of DNA repair. The DNA mismatch repair (MMR) system, which is highly conserved from prokaryotes to eukaryotes, plays an important role in maintaining genome stability by correcting base-base mismatches and insertion/deletion (ID) mispairs generated during DNA replication and other DNA transactions. Mismatch recognition is a critical step in MMR. Two mismatch recognition proteins, MutSα (MSH2-MSH6 heterodimer) and MutSβ (MSH2-MSH3 heterodimer), have been identified in eukaryotic cells. MutSα and MutSβ have partially overlapping functions, with MutSα recognizing primarily base-base mismatches and 1-2 nt ID mispairs …

Explorations In Homeoviscous Adaptation And Mass Spectral Analysis Of Membrane Lipids, Michael Douglas Timmons

University of Kentucky Doctoral Dissertations

The focus of this dissertation is centered on the mass spectral analysis of lipids and changes occurring in keeping with the concept of homeoviscous adaptation [1]. Homeoviscous adaptation is the process of modification of membrane lipids in response to environmental stimuli [1]. Dissertation investigations applied this concept to prokaryotic and eukaryotic organisms, and expanded the perception of environmental factors from exogenous organic solvents to intracellular environment.

The field of lipidomics deals with the analysis of phospholipid and fatty acid components of membranes the changes that occur due to environmental stimuli and their biological significance [2-6]. The high sensitivity of mass …

Streamflow Prediction Using Gis For The Kentucky River Basin, Bakkiyalakshmi Palanisamy

University of Kentucky Doctoral Dissertations

The study was aimed at developing a simple methodology for flow prediction in ungauged basins using existing data resources. For this purpose, the streamflow measurements across the Kentucky River Basin located in Kentucky, USA were obtained from United States Geological Survey (USGS) archive. The flow transferring characteristics of the subbasins of the Kentucky River Basin were obtained by combining downstream and upstream stream gauges. The flow transferring function thus derived were related to watershed, channel and flow characteristics of the subbasins by multiple regression analysis. The gauge pairs were divided into two classes of subbasins representing Upper and Lower Kentucky, …

Soil Water And Crop Growth Processes In A Farmer's Field, Susmitha Surendran Nambuthiri

University of Kentucky Doctoral Dissertations

The study was aimed to provide information on local biomass development during crop growth using ground based optical sensors and to incorporate the local crop status to a crop growth simulation model to improve understanding on inherent variability of crop field. The experiment was conducted in a farmer’s field located near Princeton in Caldwell County, Western Kentucky. Data collection on soil, crop and weather variables was carried out in the farm from 2006 December to 2008 October. During this period corn (Zea mays L.) and winter wheat (Triticum sp) were grown in the field. A 450 m …

15N Solid-State Nmr Detection Of Flavin Perturbation By H-Bonding In Models And Enzyme Active Sites, Dongtao Cui

University of Kentucky Doctoral Dissertations

Massey and Hemmerich proposed that the different reactivities displayed by different flavoenzymes could be achieved as a result of dominance of different flavin ring resonance structures in different binding sites. Thus, the FMN cofactor would engage in different reactions when it had different electronic structures. To test this proposal and understand how different protein sites could produce different flavin electronic structures, we are developing solid-state NMR as a means of characterizing the electronic state of the flavin ring, via the ¹⁵N chemical shift tensors of the ring N atoms. These provide information on the frontier orbitals. We propose that …

In Vivo Oxidative Stress In Alzheimer Disease Brain And A Mouse Model Thereof: Effects Of Lipid Asymmetry And The Single Methionine Residue Of Amyloid-Β Peptide, Miranda Lu Bader Lange

University of Kentucky Doctoral Dissertations

Studies presented in this dissertation were conducted to gain more insight into the role of phospholipid asymmetry and amyloid-β (Aβ)-induced oxidative stress in brain of subjects with amnestic mild cognitive impairment (aMCI) and Alzheimer disease (AD). AD is a largely sporadic, age-associated neurodegenerative disorder clinically characterized by the vast, progressive loss of memory and cognition commonly in populations over the age of ~65 years, with the exception of those with familial AD, which develop AD symptoms as early as ~30 years-old. Neuropathologically, both AD and FAD can be characterized by synapse and neuronal cell loss in conjunction with accumulation of …

Characterization Of Soil Carbon Stabilization In Long-Term Row-Cropped Agro-Ecosystems, Soraya Patricia Alvarado-Ochoa

University of Kentucky Doctoral Dissertations

Soil organic matter (SOM) is a dynamic soil property, sensitive and responsive to many factors. The possibility of increasing soil carbon (C) sequestration by changing land use and management practices has been of great interest recently due to concerns with global changes in the atmospheric carbon dioxide (CO₂) balance. Nonetheless, as a result of the complex dynamics of SOM, there is still the need for SOM characterization procedures capable of monitoring SOM stabilization, taking into account all the factors involved.

This study characterized SOM stabilization as affected by management practices in three long-term field experiments, considering physical, chemical …

Colloid Mediated Transport Of Heavy Metals In Soils Following Reclamation With And Without Biosolid Application, Jarrod O. Miller

University of Kentucky Doctoral Dissertations

Soils disturbed by strip mining practices may have increased colloid loads moving to groundwater resources, also enhancing the transport of contaminants into our water resources. We hypothesize that contaminant transport within soils following mining is enhanced by colloid mobility. Two sites were chosen for this study, a 30-year old reclaimed strip mine in southwest Virginia and a recently mined area from eastern Kentucky. Intact reclaimed soil monoliths were retrieved from sandstone derived soils in southwestern Virginia. Reclaimed monoliths from eastern Kentucky were recreated in the lab. Intact undisturbed (native) soil monoliths representing the soils before mining were also sampled for …