Life Sciences Commons^™

Characterization Of Arabidopsis Eukaryotic Translation Initiation Factor 2Α (Eif2Α) Mutants, Mark Edens

Masters Theses

Plants are stationary organisms that are charged with overcoming a multitude of biotic and abiotic stresses. The eukaryotic translation initiation factor 2 (eIF2) is responsible for charging the P-site of the forming 80S ribosome with the initiator methionyl-tRNA. Thus, eIF2 is a protein of utmost consequence in the growth and development of organisms. The eIF2 protein is also a mediator of global translational regulation in the eukaryotic integrated stress response (ISR), where the α-subunit is phosphorylated by protein kinases, such as the kinase GCN2. Here, five eIF2α allele mutants in Arabidopsis were found to be deleterious to plant growth, development, …

The Application Of Microbial Source Tracking To Aid In Site Prioritization For Remediation In Lower Michigan, John J. Hart

Masters Theses

Non-point source fecal pollution is a threat to both the environment and public health. Climate change, aging infrastructure, and intensified agricultural practices are predicted to accentuate this issue. In Michigan, due to the high instance of aging infrastructure and intensified agriculture, non-point source fecal pollution has caused many waterbodies to exceed the state standards posing a risk to recreational activities and source water. Due to this threat, there is an increased effort to identify and remediate these sources. My study focused on improving the identification of non-point source fecal pollution through a combination of culture-based and molecular fecal indicator bacteria …

Influence Of Physical Variability Of Highly Weathered Sedimentary Rock On Nitrate In Area 3 Of The Enigma Field Research Site At Y-12, Erin Kelly

Masters Theses

Uranium processing and waste storage in unlined waste ponds leached contaminants into the groundwater at Y-12, Oak Ridge, Tennessee, from the 1950s to 1980s. Groundwater wells near the S-3 ponds have had the highest nitrate concentrations of groundwater anywhere in the world (>10,000 mg/L). For reference, the maximum contaminant level for nitrate in drinking water set by the U.S. Environmental Protection Agency is 10 mg/L. Since 2012, the ENIGMA (Ecosystems and Networks Integrated with Genes and Molecular Assemblies) group has been characterizing, monitoring, and conducting field experiments to understand the interactions between contaminants, microbes, and the subsurface. The goals …

The Flavoprotein Rcla Is A Hypothiocyanous Acid Reductase, Irina Chapman

Masters Theses

Reactive chlorine species (RCS) - hypochlorous acid (HOCl), hypothiocyanous acid (HOSCN), and chloramines - are powerful antimicrobial oxidants generated by the innate immune system. Bacteria developed defense mechanisms against RCS, which are highly toxic for invading pathogens. One such defense system involves RclA, an enzyme from the flavin disulfide reductase family, which is massively upregulated upon exposure of Escherichia coli to RCS. Recent research suggests that RclA functions as a Cu(II) reductase to provide resistance against RCS. However, the exact mechanism used by RclA was subject to debate. In this study, we demonstrate that RclA is unlikely to function as …

Development Of Fluorescence Microscopy Approaches To Study Subcellular Protein Transport And Enzymatic Activity, Anchal Singh

Masters Theses

Understanding the subcellular localization of proteins and their activity is important in understanding their normal function in eukaryotic cells. Fluorescence cellular imaging techniques can selectively and sensitively visualize subcellular biochemistry. Using this approach, two different methods were employed in this thesis. The first focused on studying protein import into peroxisome and the other on monitoring the activity of an endoplasmic reticulum (ER)-localized enzyme, human carboxylesterase 1 (CES1).

Peroxisomes are mainly known as the center for long chain fatty acid b-oxidation as well as the production and detoxification of hydrogen peroxide. Proteins which are needed in the peroxisomes are encoded in …

Xylan-Derived Carbon Nutrient Acquisition By Caulobacter Crescentus, Rhoda Ekuama Inkoom

Masters Theses

Bacterial decomposition of xylan and uptake of xylan-derived carbon compounds require a combination of enzymes, proteins, and complex molecular mechanisms. Bacteria have evolved molecular mechanisms for its utilization depending on its environment and source of xylan. Caulobacter crescentus, a Gram-negative bacterium which lives in an aquatic oligotrophic environment, exhibits asymmetric division which is an adaptive feature for its nutrient poor habitat. Although aromatic hydrocarbon and cellulose-derived mono-, di- and oligosaccharide utilization has been reported in literature, its xylan-derived carbon compound utilization is yet to be characterized. In this study, C. crescentus utilizes xylan-derived monomers arabinose, xylose and acetate as …

The Impact Of Microbial Experience On The Murine Innate Immune Response, Cody Thomas Morrison

Masters Theses

The hygiene hypothesis predicts that certain environmental factors shape overall immune system function in animals and humans. While current specific pathogen free (SPF) mouse models are invaluable for studying the immune system, they have limitations for comparison with humans who have microbial exposures throughout their lifetimes. Several studies have shown that the composition of the immune system of SPF mice more closely resembles that of newborns, whereas the immune system from mice exposed to microbial pathogens more closely reflect adult immunity. In this study we have established a model using traditional SPF mice (“clean mice”) and SPF mice that were …

Seasonal Variation In Mitochondrial Bioenergetics Of The Bluegill Sunfish, Lepomis Macrochirus, From A Shallow Midwest River, Derick Isaac Lamptey

Masters Theses

As average global temperature increase, the frequency and magnitude of extreme temperatures in shallow aquatic ecosystems are more ubiquitous. In order to understand how these changing thermal regimes affect aquatic ectotherms, it is essential to develop studies evaluating the response of ectotherms to seasonal fluctuating thermal regimes. Previous studies on fluctuating temperature regimes have reported an increased physiological stress leading to morphological, behavioral and biochemical adaptations. From the latter, the adaptive capacity and seasonal performance associated with optimal function of the oxidative phosphorylation system (OXPHOS) are key for species persistence. However, studies on this matter are scarce. This study explores …

Investigation Of Glutamate Carboxypeptidase Ii Roles In The Folate Cycle Dependent Reproduction And Development Of Caenorhabditis Elegans, Jessica M. Derham

Masters Theses

Glutamate Carboxypeptidase II (GCPII) is a transmembrane, zinc metallopeptidase that is expressed in a wide range of organisms, including roundworms, mice, and humans. In humans, GCPII is primarily expressed in the prostate, kidneys, small intestine, and central nervous system. Within the small intestine, the expected function of GCPII is to aid in the absorption of dietary folate from the intestinal lumen. GCPII cleaves excess glutamates from folate to yield monoglutamated folate which is then readily transported into the enterocyte. Folate can then be used through the one carbon metabolic cycle for the synthesis of nucleotides, conversion of homocysteine to methionine, …

Therapeutic Dna: Delivery And As A Delivery Vehicle, Natalie J. Holl

Masters Theses

"A review of gene delivery methods and gene editing methods, as well as original research utilizing DNA as a delivery vehicle is presented in the following thesis. Thousands of diseases have been linked to genes. Gene therapy, either delivering therapeutic genes or editing DNA bases, has arisen as a treatment option with the potential to cure diseases, rather than just ease symptoms. Genes and editing tools need to be delivered to cells for these therapies to be effective and many techniques have been developed to address the issue of delivery. Nonviral and viral methods have been used to deliver nucleic …

Xylan Metabolism By Caulobacter Crescentus, Viet Bui

Masters Theses

Bacteria present unique opportunities to explore the molecular mechanisms of nutrient acquisition processes at single cell level under various experimental conditions. Caulobacter crescentus is a gram-negative oligotrophic environmental bacterium with adaptive physiological responses, such as slow growth or complete arrest of cell cycle, presence of sessile and motile progenies with the sessile cells growing stalk to enhance nutrient uptake for growth in low-nutrient conditions. Details of plant polysaccharide-derived carbon compound, such as cellobiose, maltose, and xylose utilization by C. crescentus can be found but molecular details or mechanisms of polysaccharide utilization, such as xylan, has not been reported. Database analyses …

Comparative Toxicology Of Nio And Ni(Oh)₂ Nanoparticles, Melissa Hope Cambre

Masters Theses

“Understanding the potential toxicity of nanoparticles (NPs) is important to ensure that these new products do not impose harmful effects to human and environmental health. Paper I is a literature review in which we discuss characteristics of nanomaterials, with an emphasis on transition metal oxide nanoparticles that influence cytotoxicity. Identification of those properties may lead to the design of more efficient and safer nanosized products for various industrial purposes and provide guidance for assessment of human and environmental health risk. We then investigate biochemical and molecular mechanisms of cytotoxicity that include oxidative stress-induced cellular events and alteration of the pathways …

Comparative Chemical Characterization Of Lunasin-Enriched Preparations And Modifications Of The Inflammasomes In Vitro, Samuel James Price

Masters Theses

Soybean (Glycine max) is one of the most cultivated crops in the world providing the population with large amounts of protein and oil. In addition to its nutritional composition, soybean also contains biologically active compounds with potential health-promoting properties. The presence of these bioactives may be responsible for the lower incidence of chronic diseases in populations that consume a significant portion of soybeans in their diet. One group of soybeanderived bioactives are bioactive peptides and proteins including lunasin, Bowman-Birk inhibitor (BBI) and Kunitz-type trypsin inhibitor (KTI). The overall objective of this research was to develop a method of …

Effects Of Asiatic Acid On Neurite Outgrowth In Neuro-2a Cells, Aishah Asiri

Masters Theses

Recently, medicinal plants from ancient Ayurvedic medicine have provided clues to the discovery of novel therapeutics for various diseases. In Ayurvedic medicine, a common Indian plant, Centella asiatica is highly regarded as a "rasayana" or nerve tonic. The Centella extract is used to ward off age-related dementia and to increase memory and intelligence. The mechanism by which Centella improves memory and learning and reduces the risk of dementia is unclear.

We recently tested the effects of asiatic acid, the main active component of Centella, on neuronal growth. We hypothesized that asiatic acid will promote neuronal growth and neurite network …

Switchgrass Extractives Have Potential As A Value-Added Antimicrobial Against Plant Pathogens And Foodborne Pathogens, Alexander Ian Bruce

Masters Theses

Panicum virgatum (switchgrass), a perennial grass native to North America, is a leading biomass feedstock candidate for the manufacture of cellulosic ethanol. Switchgrass is considered a viable option for biofuel production due to its cheap production cost and ability to grow on marginal land. Biofuel derived from switchgrass has been shown to be very energy efficient, producing 540% more renewable energy versus nonrenewable energy expended. Switchgrass-derived biofuel is also estimated to have greenhouse gas emissions that are 94% lower than emissions from gasoline (Schmer et al 2008). Biofuels are created through biochemical processes that utilize various enzymes and microorganisms for …

Lignin Maximization: Analyzing The Impact Of Different Feedstocks And Feedstock Ratios Using Organosolv Fractionation, Marc Banholzer

Masters Theses

Over-exploitation of fossil fuels coupled with increasing pressure to reduce carbon emissions are prompting a transition from conventional petrochemical feedstocks to sustainable and renewable sourced carbon. The use of lignocellulosic biomass as a feedstock for integrated biorefining is of current high interest, as separation into its component parts affords process streams of cellulose, hemicellulose and lignin, each of which can serve as a starting point for the production of biobased chemicals and fuels. Given the large number of potential sources of lignocellulosic feedstocks, the biorefinery will need to adapt to the supplies available over a normal growing season. Of particular …

Effect Of Varying Rumen Degradable And Undegradable Protein On Milk Production And Nitrogen Efficiency In Lactating Dairy Cows Under Summer Conditions, Jeffrey D. Kaufman

Masters Theses

The objective is to determine the effect of reducing nitrogen input through feeding low rumen degradable protein (RDP) and rumen undegradable protein (RUP) proportions on milk production, nitrogen efficiency and metabolism in heat-stressed cows. Forty-eight mid-lactating, Holstein cows were assigned to treatments using a randomized block design in a 2x2 factorial arrangement of treatments (n = 12/treatment). Treatments included two levels of RDP (10 and 8%) and two levels of RUP (8 and 6%). From d 1 to 21, a common diet (10% RDP-8% RUP) was fed to cows followed with their respective treatment diets fed from d 22 to …

Expression Of Insulin Responsive Genes In Insulin Resistant Conditions, And The Effect Of Selenium On Gene Expression, David L. Ruff

Masters Theses

Chronically high blood glucose levels lead to many problems, such as insulin resistance, the hallmark of Type II diabetes. Increased flux through the hexosamine biosynthesis pathway is one mechanism by which high glucose as well as glucosamine has been shown to induce insulin resistance. This study tests the effects of glucosamine induced insulin resistance on insulin regulation of the metabolic genes glucose-6-phosphate dehydrogenase (G6PDH) and fatty acid synthase (FAS) as well as insulin responsive proteins tribbles homolog (TRIB3) and sterol regulatory element binding protein (SERBP-1c) 1c.

Selenium, a micronutrient has been shown to be an effective insulin mimetic in Type …

Conformational Dynamics Of Cytochrome P450cam Upon Ligand Binding, Ana Virginia Bernal Gomez

Masters Theses

Cytochrome P450s are a superfamily of metalloenzymes that are responsible for the monooxygenation of their hydrophobic substrates. P450’s retain the same general structural scaffold, however are able to bind promiscuously to substrates with distinct physico-chemical properties. It is believed that they possess considerable flexibility in the substrate binding regions as well as their active site to accomplish the diverse binding and catalytic chemistry with high regio- and stereo-specificity. In order to investigate the conformational dynamics inherent in these enzymes, especially in context of binding different ligands, we carried out amide proton exchange studies via NMR spectroscopy on a model P450 …

Comparison Of Activation Enthalpies For Aminoglycoside Modification Reactions, Brittany Sterling Soto

Masters Theses

At highly elevated temperatures, many biological reactions can proceed spontaneously from the ground state to the transition state. However, due to the long half-life of these reactions, catalysts are required to catalyze these reactions at modern day temperatures by lowering the activation energy. Wolfenden et al. has previously shown that catalysts enhance the rate of the reaction by reducing the enthalpy of activation. Therefore, the activation energies have been determined for three aminoglycoside modifying enzymes, APH(3’)-IIIa, AAC(3)-IIIb, and AAC(3)-VIa, to determine whether these three enzymes distinguish between the two classes of aminoglycoside antibiotics by reducing the enthalpy of activation during …

Manipulating Adipose Tissue Fatty Acid Oxidation To Reduce Fatness In Broiler Chickens, Tania Emmanuelle Torchon

Masters Theses

Compared to rodents, broiler chickens, those reared for meat, are an attractive model for studies of adipose biology, and obesity development in children. The broiler chicken lacks the gene for uncoupling protein 1, the hallmark for brown adipose tissue making them a useful model to study lipid metabolism in white adipocytes. Two studies were performed to investigate if white adipose tissue had the metabolic ability for fatty acid oxidation (FAO), and to investigate the effects of dietary fatty acids on abdominal fat development of young broiler chickens as a model for childhood obesity. In study one, chickens were fasted for …

Localization Of Chemoreceptors In Azospirillum Brasilense., Anastasia Aksenova

Masters Theses

In order to ensure their survival, bacteria must sense and adapt to a variety of environmental signals. Motile bacteria are able to orient their movement in a chemical gradient by chemotaxis. During chemotaxis, environmental signals are detected by chemotaxis receptors and are propagated via a signal transduction cascade to affect bacterial motility. In a model organism Escherichia coli, chemotaxis receptors, also called MCPs (for methyl-accepting chemotaxis proteins) sense changes in concentration gradients by making temporal comparisons about the chemical composition of their surroundings. Decreased attractant concentration or increased repellant concentration results in conformational changes in the MCPs that culminate …

Value-Added Lignin Based Carbon Fiber From Organosolv Fractionation Of Poplar And Switchgrass, Andreas Attwenger

Masters Theses

Carbon fibers have unique properties that include high strength, low density and excellent chemical and thermal resistance. However, they have a low level of utilization because of their high price; typically around $30/kg for an entry level polyacrylonitrile (PAN) based carbon fiber. Low-cost carbon fibers derived from lignin are currently being investigated at the University of Tennessee, because using lignin as a precursor could significantly reduce production costs. Lignins obtained from the pulp and paper and the emerging biofuel industries have the potential to be used for carbon fiber production, however, they are typically unsuitable because of the high levels …

Examining The Functional Consequences Of The Flexibility Of Aminoglycoside Phosphotransferase (3’)-Iiia, Katelyn Dawn Rosendall

Masters Theses

The use of aminoglycoside antibiotics began in 1940 with the discovery of streptomycin. The overuse and misuse of antibiotics has resulted in prevalent cases of antibiotic resistance. The most common source of aminoglycoside resistance is the presence of enzymes that covalently modify the antibiotics at specific locations. One such enzyme, APH(3′)-IIIa [the aminoglycoside phosphotransferase three prime three a] conveys resistance by transferring the γ-phosphate [gamma phosphate] from ATP [adenosine triphosphate] onto the 3′ [three prime] carbon of the aminoglycoside antibiotic sugar ring. APH(3′)-IIIa has been shown to be flexible in solution and this flexibility is proposed to be responsible for …

Engineering Of Photosystem I Attachment To Titanium Oxide Nanostructures Via Ferredoxin-Fusion Proteins, Tuo Zhu

Masters Theses

Ferredoxins (Fds) are iron-sulfur proteins that mediate electron transfer in a range of metabolic reactions. In the thylakoid membrane of photosynthetic organisms, Fd facilitates electron transfer from the stromal surface of photosystem I (PSI) to the ferredoxin Nicotinamide adenine dinucleotide phosphate oxidoreductase (FNR), which requires that Fd is capable of docking and transferring electrons between these two complexes. In applied photosynthesis, many efforts have been devoted towards re-directing these electrons into either a hydrogen-evolving catalyst or an electron-conducting semiconductor. In this study, the electrons from the PSI complex are directed to a titanium oxide (TiO₂) electrode, and Fd …

Determination Of Structural Stabilities Of Metal-Free Porphyrin Cytochrome C, Zinc Cytochrome C, And Cobalt Cytochrome C, Usha K. Tottempudi

Masters Theses

No abstract provided.

Design And Synthesis Of Boronic Acid-Based Sensors For Microarray Analysis And Fret-Detection To Study Carbohydrates, Yilin Wang

Masters Theses

Carbohydrates play significant roles in various biological and pathological processes such as cancer metastasis and inflammation. They participate in various signaling pathways and are able to bind to a litany of biological receptors such as proteins and viruses. Biologists have known for decades that the structure of glycans changes with the onset of cancer and inflammation. Due to these important activities, carbohydrate sensing has long been a focus of research. One example of such a carbohydrate sensor is boronic acid-based sensors. Boronic acid-based sensors are of particular interest due to their selectivity for 1,2- or 1,3-diols. A variety of techniques …

Angiotensinogen Gene Silencing Reduces Lipid Accumulation And Inflammation In Cultured 3t3-L1 Adipocytes, Wenting Xin Carroll

Masters Theses

Obesity is characterized by metabolic complications which are related to several life-threatening diseases. Dysregulated inflammatory adipokines secretion from adipose tissue is believed as the major contributor to obesity-associated local and systemic inflammation, insulin resistance, and other metabolic dysfunctions.

Numerous studies in our lab and others pointed to the role of local adipose tissue renin-angiotensin system (RAS) in the pathogenesis of obesity, inflammation and insulin resistance. We hypothesized that adipocytes-derived angiotensinogen (Agt) played a critical role in adipogenesis and/or lipogenesis as well as adipose inflammation. To test, we established 3T3-L1 preadipocytes stably transfected with Agt-shRNA or scrambled sequence (Sc-shRNA). Transfected preadipocytes …

Elucidating The Effect Of Silver On Ethylene Signaling In Arabidopsis Thaliana, Brittany Kathleen Mcdaniel

Masters Theses

Ethylene, a gaseous plant hormone, is involved in numerous plant developmental processes such as seed germination, senescence, and fruit ripening. In Arabidopsis thaliana, ethylene is perceived by a family of five membrane-bound receptors, which upon binding ethylene trigger downstream effects. At the receptor level, it is known that the coordination of a copper ion is necessary for ethylene to bind, resulting in a conformational change of the receptor and the initiation of the ethylene signal transduction pathway. Interestingly, silver ions are also able to support binding of ethylene but ethylene responses are blocked in the presence of silver. When …

Capturing More Light: Phycobilisome Characterization For Increased Hydrogen Production Efficiency, Paul Abraham Willard

Masters Theses

Alternative energy and biofuels are a growing area of research. The demand for more and clean energy is ever increasing, but the current technology is inefficient, expensive, and incapable of meeting the demands of the current market. Hydrogen is a potential future fuel, as it is both clean and renewable, but its formation through conventional means is costly and inefficient. Photosynthesis can be utilized for the formation of hydrogen, which can then serve as a convenient and renewable biofuel. Photosynthetic hydrogen evolution is observed in vitro, but the current photosystem design is not very versatile and optimized to use …