Digital Commons Network^™

Computational Design And Production Of Levansucrase With Increased Thermostability, Methus Klaewkla

Chulalongkorn University Theses and Dissertations (Chula ETD)

Levansucrase is a carbohydrate-modifying enzyme that catalyzes hydrolysis of sucrose and transfructosylation of a fructosyl unit of sucrose to produce levan and levan-type fructooligosaccharides (LFOs). Due to their various beneficial properties such as low viscosity and non-toxicity. Levan and LFOs have potential applications in many industries such as prebiotics in food industries and anti-tumor agents in pharmaceutical industries. Previous study found that the Y246S mutant of Bacillus licheniformis RN-01 levansucrase (Oligosaccharide Producing Levansucrase, OPL) could effectively produce LFOs, but its thermostability is limited at high temperature. In this study, molecular dynamics (MD) and computational protein design were used to design …

Effects Of Aquatic Acidification On Calcium Uptake In White River Shrimp Litopenaeus Setiferus Gills, Maria-Flora Jacobs

UNF Graduate Theses and Dissertations

Previous research regarding aquatic acidification has examined the protonation of the carbonate and does not consider calcium to be a limiting factor. This is the first study to suggest that pH may affect the uptake of calcium in crustacean gills. This project describes ion transport mechanisms present in the cell membranes of white river shrimp Litopenaeus setiferus gill epithelium, and the effects of pH on the uptake of calcium by these means. Partially purified membrane vesicles (PPMV) of shrimp gills were prepared through a homogenization process that has been used previously to define ion transport in crab and lobster gill …

The Relationship Between Seasonal Breeding And Deiodinase Expression In The Green Anole Lizard, Hyejoo Kang

All Graduate Theses, Dissertations, and Other Capstone Projects

Reproductive physiology and behavior is mainly regulated by the hypothalamus-pituitary-gonad (HPG) axis. Interestingly, abnormal levels of thyroid hormone (TH) results in the delayed onset of puberty and affects gonadal function of adults by altering HPG axis activity. Seasonally breeding animals undergo drastic hormonal and behavioral changes between breeding and non-breeding seasons. Green anole lizards (Anolis carolinensis), similar to other seasonally breeding animals, have increased sex steroid hormones, larger gonads, upregulated gonadal steroidogenic acute regulatory protein (StAR) mRNA and increased reproductive behavior during the breeding compared to non-breeding seasons. Relatively less is known regarding the regulation of gonadal TH in seasonal …

Mutations Of Fus Cause Aggregation Of Rna Binding Proteins, Disruptions In Protein Synthesis, And Dysregulation Of Nonsense Mediated Decay, Marisa Elizabeth Kamelgarn

Theses and Dissertations--Toxicology and Cancer Biology

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by motor neuron death and subsequent muscle atrophy. Approximately 15% of ALS cases are inheritable, and mutations in the Fused in Sarcoma (FUS) gene contribute to approximately 5% of these cases, as well as about 2% of sporadic cases. FUS performs a diverse set of cellular functions, including being a major regulator of RNA metabolism. FUS undergoes liquid- liquid phase transition in vitro, allowing for its participation in stress granules and RNA transport granules. Phase transition also contributes to the formation of cytoplasmic inclusions found in the …

Molecular Level Comparison Of Water Extractives Of Maple And Oak With Negative And Positive Ion Esi Ft-Icr Mass Spectrometry, Zhongqi He, Rachel L. Sleighter, Patrick G. Hatcher, Shasha Liu, Fengchang Wu, Haixuan Zou, O. Modesto Olanya

Chemistry & Biochemistry Faculty Publications

Soluble extractives in wood function to protect living trees from destructive agents and also contribute to wood color and fragrance. Some extractive components have biological activities with medical applications. They also play important roles in wood processing and related applications. To increase the knowledge of wood chemistry, maple and oak were extracted by water. Ultraviolet/visible (UV/vis) spectroscopy indicated the presence of a phenolic compound, resorcinol, in maple extractives having higher molecular mass and more aromatic components than oak extractives. Negative and positive electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI FT-ICR-MS) identified thousands of formulas in the two …

Island Invasion: The Silent Crisis In Hawaii, Sophia Janssen

Pomona Senior Theses

Keeping out invasive species may, upon first review, seem like a trivial environmental cry from ecologists and deep environmentalists; a belated wish to return to an undeveloped world where nature was pristine. However invasive species create problems that impact all of us and can have far more severe consequences than changing a stunning landscape. These problems are heightened in islands like Hawaii, where the fragile ecosystems have developed over centuries of evolution and adaptation. The introduction of a disease-carrying mosquito can put the people of Hawaii at risk to many vector-born illnesses and create an epidemic, taking human life. The …

Condensation And Polymerization Explain The Humification Of Lignin Into Aliphatic And Aromatic Structures In Soil, Patrick G. Hatcher, Hongmei Chen, Seyyedhadi Khatami, Derek C. Waggoner

Chemistry & Biochemistry Faculty Publications

Soil organic matter (SOM) constitutes a global reservoir of carbon that is more than twice that of either atmospheric carbon or aquatic carbon; however, the manner in which it forms from degraded plant biomass is poorly understood. Some have recently questioned whether plant biomass is involved directly in SOM formation and suggest that it is microbial carbon that constitutes the main source of stable SOM. Such a view implies that above and below ground plant biomass is rapidly decomposed and mineralized. This view contrasts significantly with traditional ones that involve the transformation of plant biomass to recalcitrant humic materials fueled …

Treatment Of Plastic Wastes Using Plasma Gasification Technology, Zachary Homolka

Honors Program: Senior Projects (Public)

Plasma gasification (PG) complements traditional recycling when applied to contaminated or mixed plastics. Without PG these plastics cost recyclers more to process than they are worth on the market, and sometimes they are landfilled or incinerated instead of being recycled. Plasma gasification can take plastic not suitable for traditional recycling and break it down into high-quality syngas for use in electricity generation, chemical manufacturing, or hydrogen production. The technology can be implemented without changing the behavior of consumers, which is a major advantage over attempting to decrease contamination or reduce use of low-value plastic. Due to high capital requirements and …

Amino Acids Profiling For The Diagnosis Of Metabolic Disorders, Yana Sandlers

Chemistry Faculty Publications

Inborn errors of metabolism (IEM) represent a group of inherited diseases in which genetic defect leads to the block on a metabolic pathway, resulting in a single enzyme dysfunction. As a downstream consequence of the residual or full loss of the enzymatic activity, there is an accumulation of toxic metabolites in the proximity of the metabolic block and/or a deficiency of an essential metabolic product which leads to the clinical presentation of the disease. While individually IEMs are rare, a collectively estimated incidence of metabolic inherited disorders is 1:800. The genetic basis of IEMs can involve abnormalities such as point …

Unintended Consequences Of Dna Analysis Delays In North Carolina, Pamela Cook Woodard

Walden Dissertations and Doctoral Studies

The processing of DNA recovered from felony crime scenes often causes delays in trials of up to 3 years, calling into question defendants' rights to a speedy trial. Using Lewin's force field analysis as the theoretical framework, the purpose of this quantitative, comparative study was to compare the processing and reporting of results related to DNA testing in 4 states. Survey data were collected from state bar members (n=137), members of a professional law organization (n=149), and members of a state DNA laboratory (n=20). The purpose of this quantitative, comparative study was to determine whether these variables (interagency communications, staff …

Toward An Enzyme-Coupled, Bioorthogonal Platform For Methyltransferases: Probing The Specificity Of Methionine Adenosyltransferases, Tyler D. Huber

Theses and Dissertations--Pharmacy

Methyl group transfer from S-adenosyl-l-methionine (AdoMet) to various substrates including DNA, proteins, and natural products (NPs), is accomplished by methyltransferases (MTs). Analogs of AdoMet, bearing an alternative S-alkyl group can be exploited, in the context of an array of wild-type MT-catalyzed reactions, to differentially alkylate DNA, proteins, and NPs. This technology provides a means to elucidate MT targets by the MT-mediated installation of chemoselective handles from AdoMet analogs to biologically relevant molecules and affords researchers a fresh route to diversify NP scaffolds by permitting the differential alkylation of chemical sites vulnerable to NP MTs that are unreactive to …

The Involvement Of Progesterone In Glial Cell Viability, Tracee L. Guthrie

Theses and Dissertations

We studied the effects of ethynodiol diacetate on the viability of hydrogen peroxide stressed glial cells. We wanted to determine the extent in which varying concentration of ethynodiol diacetate affect cell viability of cultured glial cells from hydrogen peroxide toxicity. Based on previous studies, we predicted that the viability of hydrogen peroxide-stressed cells will increase with the presence of the progesterone, ethynodiol diacetate, and that cell viability will increase with increasing concentration of ethynodiol diacetate. Glial cells of 1x10° cells/well were introduced into a 96-well plate and allowed to grow for 24 hours in an incubator at 37°C and 5% …

The Effects Of Ethynodiol Diacetate On Epinephrine-Treated Glial-Derived Cells, Jesse D. Hunt

Theses and Dissertations

Neuroprotective agents, including specific hormones, may play a role in preventing cellular damage following an ischemic event or brain injury. Astrocytes are specialized glial cells within the central nervous system that play roles in synapse development, blood flow regulation, extracellular homeostasis, synapse function, energy and metabolism (Sofroniew and Vinters, 2009). Research using astrocytes in cell viability studies can be beneficial in deepening our understanding of specific neuroprotective effects of various steroid hormones including progesterone. Progesterone has been shown to have neuroprotective capabilities in the central nervous system (Scarpin et ah, 2009). Although progesterone shows great therapeutic potential, effects of the …

The Effects Of Heat-Killed Echinostomatid Parasites On Mice Treated With Dextran Sodium Sulfate (Dss), Natalie Gooder

All Graduate Theses, Dissertations, and Other Capstone Projects

Objective: Autoimmune diseases are chronic, incurable, and affect approximately 50 million Americans. This is a strong need for better ways to treat autoimmune diseases. Parasites and parasite proteins have been observed to protect mice from symptoms of induced colitis in mice treated with dextran sodium sulfate (DSS), a model for inflammatory bowel disease (IBD). I hypothesize that non-living echinostomatid parasites, given during DSS treatment, can decrease intestinal inflammation and weight loss, providing a possible novel treatment for IBD.

Methods: A range of DSS concentrations (0.5%-3%) were delivered to female C57BL/6 mice in their drinking water in order to determine the …

Near-Infrared Fluorescent Probes For Sensitive Determination Of Lysosomal & Mitochondrial Ph In Live Cells, Wafa Mazi

Dissertations, Master's Theses and Master's Reports

Varied intracellular pH levels are critical for various physiological processes such as enzymatic activity, cell proliferation and apoptosis, ion transport, and muscle contraction. Cellular compartments, like lysosomes, must retain an acidic environment (pH ~ 4.5) to activate hydrolytic enzymes necessary for the breakdown of large biomolecules. Another cellular organelle, the mitochondria, provides the cell with energy and must retain an alkalis environment (pH ~ 8.0) for proper function. Substantial lysosomal and mitochondrial pH deviation is associated with cellular dysfunction and disease. Therefore, the precise detection of lysosomal and mitochondrial pH is essential to provide a better understanding of cellular physiological …

Changes On Movement Control Of Dart Throwing Under Distance And Target Weight Constraints, Carlos E. Campos, Guilherme M. Lage, Andre G.P. Andrade, Crislaine R. Couto, Suziane P. Santos, Vitor L.S. Profeta, Herbert Ugrinowitsch

Athletic Performance Research

The aim of the study was to verify the effects of dart weight and target distance on kinematic variables of the movement control of the underarm dart throwing task. Four women and one man performed the task of throwing a dart at two horizontal circular targets located at 2m (Nt) and 3m (Ft) away, with two different weights, 22g (Ld) and 44g (Hd). On the first phase of the experiment, the human volunteers performed 200 trials per day during four sessions. On the fifth day, it had 40 more trials in a pseudo random order that were recorded and analysed. …

Characterizing The Cellulose-Modifying Enzyme Bcsg From Escherichia Coli, Alexander Anderson, Joel T. Weadge

Theses and Dissertations (Comprehensive)

Microbial biofilms are communities of microorganisms that exhibit co-operative behaviour, producing a matrix of exopolysaccharide that enmeshes the community. The well-studied human pathogens Escherichia coli and Salmonella entericaproduce a biofilm matrix comprised chiefly of the biopolymer cellulose, along with amyloid protein fibers termed curli. This biofilm matrix confers surface adherence and acts as a protective barrier to disinfectants, antimicrobials, environmental stressors, and host immune responses. Pertaining to this research, the bcsEFG operon, conserved in the Enterobacteriaceae, encodes an inner membrane-spanning complex responsible for the addition of a phosphoethanolamine (pEtN) modification to microbial cellulose, essential for extracellular matrix assembly and …

Characterization Of The Red Complex Bacterial Biofilm, Alena Pratasouskaya

Theses and Dissertations (Comprehensive)

Commonly associated with severe inflammation and destruction of the tooth-supporting tissue is bacterial consortia Treponema denticola, Tannerella forsythia, and Porphyromonas gingivalis, collectively referred to as the red complex. The red complex uses the common bacterial strategy of producing and embedding itself in extracellular polymeric substance, which contributes to the recalcitrance of periodontitis and was therefore of interest in this study. The red complex static cultures were grown in combination with different chemicals in order to establish what changes accompany these chemical challenges. This research established that extracellular polymeric substance carbohydrate yields increased with time. Interestingly, carbohydrate composition …

Global Patterns And Drivers Of Ecosystem Functioning In Rivers And Riparian Zones, Scott D. Tiegs, David M. Costello, Mark W. Isken, Guy Woodward, Peter B. Mcintyre, Mark O. Gessner, Eric Chauvet, Natalie A. Griffiths, Alexander S. Flecker, Vicenç Acuña, Ricardo J. Albariño, Daniel C. Allen, Cecilia Alonso, Patricio Andino, Clay Arango, Jukka Aroviita, Marcus V. M. Barbosa, Leon A. Barmuta, Colden V. Baxter, Jose Checo Colón-Gaud

Biology: Faculty Publications

River ecosystems receive and process vast quantities of terrestrial organic carbon, the fate of which depends strongly on microbial activity. Variation in and controls of processing rates, however, are poorly characterized at the global scale. In response, we used a peer-sourced research network and a highly standardized carbon processing assay to conduct a global-scale field experiment in greater than 1000 river and riparian sites. We found that Earth’s biomes have distinct carbon processing signatures. Slow processing is evident across latitudes, whereas rapid rates are restricted to lower latitudes. Both the mean rate and variability decline with latitude, suggesting temperature constraints …

Non-Contact Trapping And Stretching Of Biological Cells Using Dual-Beam Optical Stretcher On Microfluidic Platform, Aotuo Dong, Balaadithya Uppalapati, Shariful Islam, Brandon Gibbs, Ganesan Kamatchi, Sacharia Albin, Makarand Deo

Electrical & Computer Engineering Faculty Publications

Optical stretcher is a tool in which two counter-propagating, slightly diverging, and identical laser beams are used to trap and axially stretch microparticles in the path of light. In this work, we utilized the dual-beam optical stretcher setup to trap and stretch human embryonic kidney (HEK) cells and mammalian breast cancer (MBC) cells. Experiments were performed by exposing the HEK cells to counter-propagating laser beams for 30 seconds at powers ranging from 100 mW to 561 mW. It was observed that the percentage of cell deformation increased from 16.7% at 100 mW to 40.5% at 561 mW optical power. The …

Measuring Trace Element Concentrations In Artiodactyl Cannonbones Using Portable X-Ray Fluorescence, Joshua L. Henderson

All Master's Theses

Artiodactyl bones are the most common faunal remains found in Washington prehistoric archaeology sites, but they are often too fragmented to accurately identify a family, genus, or species. Traditional faunal analysis can only organize unidentifiable bone fragments into size class, and chemical methods often require the destruction of bone samples. In this thesis research, I tested a new, nondestructive faunal analysis technique using portable X-ray fluorescence (pXRF) to measure trace element concentrations in comparative collection and archaeological bone samples. Using cannonbones from five different artiodactyl species, I collected trace element data from 50 comparative collection specimens and 18 archaeological specimens …

Investigating Potential Multi-Protein Complexes Involving Emx2, Nicole Enger

All Master's Theses

The neocortex of the mammalian brain is allocated into specialized areas during embryonic development. In mice, as well as humans, the neocortex develops four primary areas: somatosensory, auditory, visual, and motor. In the adult neocortex, the boundaries between these four areas are marked by differences in cellular architecture and gene expression patterns. However, these physical boundaries are not evident during embryonic development of this structure. Rather, the neocortex initially appears uniform across its expanse, with one exception: a handful of proteins that regulate neocortical arealization are generated in gradients across the expanse of the neocortex and control the size of …

Effects Of Hydrogen Synthesis Inhibition On Changes Of Bioproduct Contents In Cyanobacterium Synechocystis Sp. Pcc 6803, Nannaphat Sukkasam

Chulalongkorn University Theses and Dissertations (Chula ETD)

Cyanobacteria have an ability to convert carbon dioxide and solar light to bioproducts. Synechocystis sp. PCC 6803 is the laboratory model strain of cyanobacteria with known metabolism and biochemistry. NADPH is one of the essential electron carriers and coenzymes required for various cellular metabolism including the production of bioproducts. [NiFe]-Hydrogenase consumes NADPH to produce hydrogen gas which considerably the loss of metabolic energy. Thus, the elimination of [NiFe]-hydrogenase would increase NADPH level in the cell and might increase the accumulation of bioproducts. In this work, hoxY and hoxH genes (encoding the two catalysis subunits of [NiFe]-hydrogenase) were inactivated. The inactivation …

Anti-Tumor Activities Of Auricularia Polytricha Extracts In Hepatocellular Carcinoma Cell Line (Hepg2) And Breast Cancer Cell Lines (Mcf-7 And Mda-Mb-231), Sunita Nilkhet

Chulalongkorn University Theses and Dissertations (Chula ETD)

Cancer is the disease that happens to occur in a high number of new cases each year. Even the choice of treatment is developing, but the adverse effects on cancer therapeutic drugs still challenging. To investigate the antitumor activities of Auricularia polytricha (AP) mushroom extract and active compounds, human cancer cell lines such as Hepatocellcular (HepG2) and Breast cancer cell lines (MCF-7 and MDA-MB-231) were interested. The crude hexane extract of AP (APH) and crude ethanol extract of AP (APE) were prepared by maceration and Soxhlet extraction, respectively. Both APE and APH were investigated for their compound compositions by GC-MS. …

Biomass Yield In A Genetically Diverse Miscanthus Sinensis Germplasm Panel Evaluated At Five Locations Revealed Individuals With Exceptional Potential, Lindsay V. Clark, Maria S. Dwiyanti, Kossonou G. Anzoua, Joe E. Brummer, Bimal Kumar Ghimire, Katarzyna Glowacka, Megan Hall, Kweon Heo, Xiaoli Jin, Alexander E. Lipka, Junhua Peng, Toshihiko Yamada, Ji Hye Yoo, Chang Yeon Yu, Hua Zhao, Stephen P. Long, Erik J. Sacks

Department of Biochemistry: Faculty Publications

To breed improved biomass cultivars of Miscanthus ×giganteus, it will be necessary to select the highest‐yielding and best‐adapted genotypes of its parental species, Miscanthus sinensis and Miscanthus sacchariflorus. We phenotyped a diverse clonally propagated panel of 569 M. sinensis and nine natural diploid M. ×giganteus at one subtropical (Zhuji, China) and five temperate locations (Sapporo, Japan; Leamington, Ontario, Canada; Fort Collins, CO; Urbana, IL; and Chuncheon, Korea) for dry biomass yield and 14 yield‐component traits, in trials grown for 3 years. Notably, dry biomass yield of four Miscanthus accessions exceeded 80 Mg/ha in Zhuji, China, approaching …

An Arabidopsis Protoplast Isolation Method Reduces Cytosolic Acidification And Activation Of The Chloroplast Stress Sensor Sensitive To Freezing 2, Allison C. Barnes, Christian G. Elowsky, Rebecca Roston

Department of Biochemistry: Faculty Publications

Chloroplasts adapt to freezing and other abiotic stresses in part by modifying their membranes. One key-remodeling enzyme is SENSITIVE TO FREEZING2 (SFR2). SFR2 is unusual because it does not respond to initial cold stress or cold acclimation, instead it responds during freezing conditions in Arabidopsis. This response has been shown to be sensitive to cytosolic acidification. The unique lipid products of SFR2 have also been detected in response to non-freezing stresses, but what causes SFR2 to respond in these stresses is unknown. Here, we investigate protoplast isolation as a representative of wounding stress. We show that SFR2 oligogalactolipid products accumulate …

Ehdv-2 Infection Prevalence Varies In Culicoides Sonorensis After Feeding On Infected White-Tailed Deer Over The Course Of Viremia, Sandra Y. Mendiola, Mary K. Mills, Elin Maki, Barbara S. Drolet, William C. Wilson, Roy D. Berghaus, David E. Stallknecht, Jonathan Breitenbach, D. Scott Mcvey, Mark G. Ruder

School of Veterinary and Biomedical Sciences: Faculty Publications

Epizootic hemorrhagic disease viruses (EHDVs) are arboviral pathogens of white-tailed deer and other wild and domestic ruminants in North America. Transmitted by various species of Culicoides, EHDVs circulate wherever competent vectors and susceptible ruminant host populations co-exist. The impact of variation in the level and duration of EHDV viremia in white-tailed deer (Odocoileus virginianus) on Culicoides infection prevalence is not well characterized. Here we examined how infection prevalence in a confirmed North American vector of EHDV-2 (Culicoides sonorensis) varies in response to fluctuations in deer viremia. To accomplish this, five white-tailed deer were experimentally infected with EHDV-2 and colonized C. …

Determining The Role Of Nudt7 In The Regulation Of Cellular Coa Levels And Metabolism, Stephanie Anne Shumar

Graduate Theses, Dissertations, and Problem Reports (ETD)

Coenzyme A (CoA) is an essential cofactor required for hundreds of metabolic processes. Because it is such a critical cofactor, CoA levels are tightly regulated. In the fasted state and in diabetic mice, the concentration of CoA increases dramatically in the liver. This phenotype is associated with constitutively low CoA degradation, a process that is emerging as a potentially important mechanism for CoA regulation. Nudt7 and Nudt19 are two mammalian peroxisomal enzymes with CoA-degrading activity, which are highly expressed in the liver and kidney, respectively. Limited information is available on the biochemistry of Nudt7 and Nudt19; the structural basis for …

Exploration Of Thiol-Michael Addition Bioconjugation To Extend Polymers Of A Protein-Based Hydrogel, Elaine Wallin Smith

Honors Theses

Hydrogels have been explored for many biomedical applications, including targeted, in situ drug delivery to avoid the negative side effects associated with systemic delivery. In our work, we are exploiting the high affinity, calcium-dependent binding between calmodulin and its target peptides to create a biomaterial for in situ, extracellular drug delivery. Genes were engineered to make two protomers, Calmodulin Collagen-Like Protein (CCLP) and Peptide Collagen-Like Protein (PCLP), that will spontaneously self-assemble in situ due to the high Ca2+ concentration in the extracellular space and provide tunable, targeted drug delivery in a biocompatible hydrogel. One important factor that would dictate the …

Preparation Of A Perylenebisimide Acceptor With Serinol And Nonadecyl-Swallowtail Imides For Use In Molecular Rectification, Tarrah Frederick

Honors Theses

Several attempts were made to add 2-amino-1,3-propanediol to the acceptor perylene monoanhydride (PMA). The theoretical product, N-(10-nonadecyl)-N'-(1,3-dihydroxypropan-2-yl)perylene-3,4,9,10-bis(dicarboximide), is intended for use in unimolecular rectification because the perylenebisimide core is classified as a good acceptor due to its high electron affinity. Rectification usually utilizes donor-σ-acceptor molecules, whose localized orbitals permit electron transfer when placed between metal electrodes of asymmetric voltages. However, according to an “asymmetric rectification” analysis, an electron can pass from one electrode, through the LUMO of the acceptor, and then to the other electrode when said electrodes are held at certain voltages, and result in rectification if the acceptor …