Molecular Biology Commons^™

Fabricating Cost-Effective Nanostructures For Biomedical Applications, Erden Ertorer

Electronic Thesis and Dissertation Repository

In this thesis we described inexpensive alternatives to fabricate nanostructures on planar substrates and provided example applications to discuss the efficiency of fabricated nanostructures.

The first method we described is forming large area systematically changing multi-shape nanoscale structures on a chip by laser interference lithography. We analyzed the fabricated structures at different substrate positions with respect to exposure time, exposure angle and associated light intensity profile. We presented experimental details related to the fabrication of symmetric and biaxial periodic nanostructures on photoresist, silicon surfaces, and ion-milled glass substrates. Behavior of osteoblasts and osteoclasts on the nanostructures was investigated. These results …

Application Of Computational Molecular Biophysics To Problems In Bacterial Chemotaxis, Davi Ortega

Doctoral Dissertations

The combination of physics, biology, chemistry, and computer science constitutes the promising field of computational molecular biophysics. This field studies the molecular properties of DNA, protein lipids and biomolecules using computational methods. For this dissertation, I approached four problems involving the chemotaxis pathway, the set of proteins that function as the navigation system of bacteria and lower eukaryotes.

In the first chapter, I used a special-purpose machine for molecular dynamics simulations, Anton, to simulate the signaling domain of the chemoreceptor in different signaling states for a total of 6 microseconds. Among other findings, this study provides enough evidence to propose …

Oligodeoxynucleotide Synthesis Using Protecting Groups And A Linker Cleavable Under Non-Nucleophilic Conditions, Xi Lin

Dissertations, Master's Theses and Master's Reports - Open

Oligodeoxynucleotides (ODNs) containing latent electrophilic groups can be highly useful in antisense drug development and many other applications such as chemical biology and medicine, where covalent cross-linking of ODNs with mRNA, protein and ODN is required. However, such ODN analogues cannot be synthesized using traditional technologies due to the strongly nucleophilic conditions used in traditional deprotection/cleavage process.

To solve this long lasting and highly challenging problem in nucleic acid chemistry, I used the 1,3-dithian-2-yl-methoxycarbonyl (Dmoc) function to protect the exo-amino groups on the nucleobases dA, dC and dG, and to design the linker between the nascent ODN and solid support. …

Refining Molecular Approaches For Bacterial Source Tracking (Bst) In Texas, Joy Truesdale

Open Access Theses & Dissertations

Bacterial source tracking (BST) is a new science that aids in identifying sources of human and animal fecal contamination of environmental water. There are currently two approaches used in BST, library-dependent and library-independent methods. The fecal bacterium E. coli is often used in library dependent methods because it indicates fecal contamination, has been used in human health risk assessments, and has water quality regulatory significance. A disadvantage of E. coli library dependent BST methods is that identification libraries of E. coli isolated from known fecal sources are needed to identify the sources of E. coli isolated from water samples. Further, …

Amalgamation Of Nucleosides And Amino Acids In Antibiotic Biosynthesis, Sandra H. Barnard

Theses and Dissertations--Pharmacy

The rapid increase in antibiotic resistance demands the identification of novel antibiotics with novel targets. One potential antibacterial target is the biosynthesis of peptidoglycan cell wall, which is both ubiquitous and necessary for bacterial survival. Both the caprazamycin-related compounds A-90289 and muraminomicin, as well as the capuramycin-related compounds A-503083 and A-102395 are potent inhibitors of the translocase I enzyme, one of the key enzymes required for cell wall biosynthesis. The caprazamycin-related compounds contain a core nonproteinogen b-hydroxy-a-amino acid referred to as 5’-C-glycyluridine (GlyU). Residing within the biosynthetic gene clusters of the aforementioned compounds is a shared open reading …

Identification Of Persistent Long Range Interactions In GA95 And GB95 Through Thermal Unfolding Simulations, Milen Redai Tesfamariam

Chemistry & Biochemistry Theses & Dissertations

For over five decades, different experiments have been performed to research how proteins attain their native three dimensional structures. However, the folding problem continues to be a puzzle in modern science. The design of two proteins that have maximal sequence identity but different folds and functions is one method that is being used to study the relationship between protein structure and amino acid sequence. In particular, mutant proteins of Streptococcus protein G, G_A and G_B, have 95% sequence identity and a 3a helix fold and β4/a fold, respectively. Molecular dynamics simulations of G_A95 …

Structural Motifs Of Novel Metallothionein Proteins, Duncan E K Sutherland

Electronic Thesis and Dissertation Repository

Metallothioneins (MT) are a family of small cysteine rich proteins, which have been implicated in toxic metal detoxification, protection against oxidative stress, and as a metallochaperone. The most well studied member of the family is the mammalian MT, which consists of two domains: a β-domain with 9 cysteine residues, which sequesters 3 Cd²⁺/Zn²⁺, and an α-domain with 11 cysteine residues, which sequesters 4 Cd²⁺/Zn²⁺. The exact functions of MT are unknown but must relate to its metalation status. Several areas that could lead to the assignment of function include 1) the determination …

Genetic And Biochemical Studies Of Human Apobec Family Of Proteins, Priyanga Wijesinghe

Wayne State University Dissertations

The AID/APOBEC family of proteins in higher vertebrates converts cytosines in DNA or RNA into uracil. These proteins have essential roles in either innate immunity or adaptive immunity. Recently, AID has also been implicated in DNA demethylation in the context of early embryogenesis in mammals. This is partly based on the reported ability of AID to deaminate 5-methyl cytosine to thymine (5mC to T). I reexamined this proposed new role of AID (5mC deamination) with two members of the APOBEC family in a novel Escherichia coli based genetic system. My results confirmed that while all three enzymes are strong cytosine …

Inhibition Of Glutamate Receptors By Constructing Bipartite Rna Aptamers, Jeffrey Hebert

Legacy Theses & Dissertations (2009 - 2024)

The relationship of excessive activity of AMPA-type glutamate receptors, and cell death, has long provided researchers a means of investigating neurodegenerative disorders, such as Parkinson's Disease (PD) and amyotrophic lateral sclerosis (ALS). Inhibitors of AMPA receptor channels, including chemical and nucleic acid molecules such as RNA aptamers, have served as potential therapeutic agents and treatment of neurodegenerative disorders. In this study, building bipartite aptamers to enhance inhibitory potency, as compared with a monomeric aptamer of AMPA receptor, is described. An enhanced potency is due, at least in part; to the proximity effect in bipartite structures or binding of a monomeric …

A Time-And-Space Parallelized Algorithm For The Cable Equation, Chuan Li

Doctoral Dissertations

Electrical propagation in excitable tissue, such as nerve fibers and heart muscle, is described by a nonlinear diffusion-reaction parabolic partial differential equation for the transmembrane voltage $V(x,t)$, known as the cable equation. This equation involves a highly nonlinear source term, representing the total ionic current across the membrane, governed by a Hodgkin-Huxley type ionic model, and requires the solution of a system of ordinary differential equations. Thus, the model consists of a PDE (in 1-, 2- or 3-dimensions) coupled to a system of ODEs, and it is very expensive to solve, especially in 2 and 3 dimensions.

In order to …

Microbial Landscapes Of Corals And Ctenophores, Camille Arian Daniels

USF Tampa Graduate Theses and Dissertations

As technology and engineering allow mankind to survey nature at finer scales, the importance of bacteria has been elucidated in their metabolic diversity, ability to transfer genetic information, involvement in biogeochemical cycling, and sheer abundance. With an individual domain of life unto themselves, this diverse group of microorganisms plays an integral role in facilitating life on land and in the oceans, and is second only to viruses in abundance on Earth. They carve niches in a wide range of environments, including those inhospitable to other life forms, and reside in concert or to the detriment of other microbes and/or hosts …

Molecular Marker Development For The Discrimination Of Atlantic And Pacific Blue Marlin (Makaira Nigricans), Laurie Sorenson

Dissertations, Theses, and Masters Projects

No abstract provided.

Human Decomposition Ecology At The University Of Tennessee Anthropology Research Facility, Franklin Edward Damann

Doctoral Dissertations

The University of Tennessee Anthropology Research Facility (ARF) is well known for its unique history as a site of human decomposition research in a natural environment. It has been integral to our understanding of the processes of human decomposition. Over the last 30 years 1,089 bodies have decomposed at this 1.28 acre facility, producing a density of 850 corpses per acre of land. This project evaluated the abiotic and biotic characteristics of the soil exposed to various levels of human decomposition in order to determine the effect on the physicochemical properties and the indigenous bacterial communities.

Specifically, 75 soil samples …

Characterization Of Metastasis-Associated Cell Surface Glycoproteins In Prostate Cancer, Lifang Yang

Theses and Dissertations in Biomedical Sciences

Prostate cancer (PCa) is a major health problem in males in the United States. Its lethality is mostly attributed to the primary tumor metastasizing to distant sites that are highly resistant to conventional therapies. Serum Prostate Specific Antigen (PSA) is the only protein biomarker used in clinic for prediction of prostate cancer recurrence following local therapies. Nonetheless, PSA lacks the ability to predict the behavior of an individual tumor in an individual patient. Therefore, development of reliable biomarkers for detection of metastatic potential in primary tumors, as well as discovery of new therapeutic targets, is in a great need for …

A Synthetic Biological Engineering Approach To Secretion- Based Recovery Of Polyhydroxyalkanoates And Other Cellular Products, Elisabeth Linton

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The costs associated with cellular product recovery commonly account for as much as 80% of the total production expense. As a specific example, significant recovery costs limit commercial use of polyhydroxyalkanoates (PHA), which comprise a class of microbially-accumulated polyesters. PHAs are biodegradable compounds that are of interest as a sustainable alternative to petrochemically-derived plastics. Secretion-based recovery of PHAs was studied to decrease PHA production costs. Type I and II secretory pathways are commonly used for the translocation of recombinant proteins out of the cytoplasm of E. coli. Proteins were targeted for translocation using four signal peptides (HlyA, TorA, GeneIII, …

Identifying Biomarkers For Resistance To Novel Cisplatin Analogues In Human Lung, Breast And Prostate Cancers, Becky Michelle Hess

UNLV Theses, Dissertations, Professional Papers, and Capstones

Cisplatin is a common therapeutic agent used in cancer treatment. Unfortunately, resistance to cisplatin in addition to severe side effects limits its use in cancer treatment. Two novel cisplatin analogues, 4DB and 4TB were shown to have varying cytotoxicity in lung, breast and prostate cancer cells. The hypothesis for this study states that the differences in 4DB and 4TB cytotoxicity among different tissue types is due to the type and efficiency of DNA repair mechanisms involved in response to these drugs.

To test the hypothesis, proteins involved in the rate limiting step of nucleotide excision repair (NER) and mismatch repair …

Characterization Of Antibiotic Resistance In Vibrio Cholerae Isolated From Ships' Ballast And Other Environmental Sources, Frank Kornegay Thomson Iii

OES Theses and Dissertations

Ships' ballast has been implicated as a vector in the dispersal of invasive species into new environments. Numerous cases have been documented for the introduction of macroorganisms via ballasting operations, but what is less known is the role of ballast water and residuals in the spread of potentially harmful microbes, especially with reference to novel genotypes. Of particular interest are the genes encoding for various forms of antibiotic resistance, many of which are carried on mobile genetic elements. A four-year sampling effort has yielded over 300 putative isolates of Vibrio cholerae from ships' ballast tanks and various environmental sources, of …

Assessing Genetic Variation Within And Among Native Populations And Hatchery Stocks Of Crassostrea Ariakensis Using Microsatellite Markers, Jie Xiao

Dissertations, Theses, and Masters Projects

Assessing genetic variation within and among native populations and hatchery stocks of Crassostrea ariakensis using microsatellite markers In response to the dramatic decline of eastern oyster (Crassostrea virginica) populations in the Chesapeake Bay, introduction of the non-native Asian oyster Crassostrea ariakensis has been proposed. Currently several hatchery stocks of C. ariakensis, derived from a few wild populations along the coast of Japan and China, are being maintained in U.S. hatcheries in the Pacific northwest and on the east coast. In recent years, as the risks of the proposed introduction are being assessed, these hatchery animals have been widely used for …

A Locus-Based Paradigm For Generating Systems Biological Inferences From Large Scale Functional Genomics Datasets, Ajish Dominic George

Legacy Theses & Dissertations (2009 - 2024)

Genomics data is growing at a exponential rate. The ability to integrate new results with existing knowledge about genomic biology is rapidly becoming the limiting factor as there no universal language with which to describe genomic functional elements. To integrate and compare new and existing genomic data, we define our basic functional unit of a genome to be a locus -- a set of positional coordinates along any genome with an arbitrary amount of functional annotations attached. The locus concept enables addressing genomic elements and annotations at any level of granularity from entire swaths of chromosomes to single base-positions. We …

The Development Of A "Genetic Eyewitness" Profiling System For Low Template Forensic Specimens: Identification Of Novel Protein, Rna, And Dna Biomarkers, Erin Hanson

Electronic Theses and Dissertations

In many criminal investigations, valuable information regarding the physical appearance of suspected perpetrators or the time and order of events that transpired are provided by eyewitness accounts. However, the information obtained from eyewitnesses is often constrained by human recollection or subjective accounts and provides a biased description of the perpetrator's appearance or an inaccurate time line of events. Additionally, in numerous situations eyewitness accounts may not be available. An increasing reliance therefore is placed on the biological evidence recovered during criminal investigations to act as a silent witness, providing unbiased and scientific information that may aid in the resolution of …

A Molecular Analysis Of Atlantic Menhaden (Brevoortia Tyrannus) Stock Structure, Abigail J. Lynch

Dissertations, Theses, and Masters Projects

Atlantic menhaden is an ecologically and economically important species along the U.S. east coast. As a filter-feeder and key prey fish, it provides a critical link between primary production, phytoplankton, and larger piscivorous predators, such as striped bass, bluefish, and weakfish. The species is also the target of one of the largest commercial fisheries in the country. Menhaden are assessed as a single, coastwide stock, and recent assessments indicate that it is not overfished. However, there is very limited population genetics data to support the assumption of a single stock. Additionally, the recent consolidation of the fishery and localization of …

Improved Constrained Global Optimization For Estimating Molecular Structure From Atomic Distances, Terri Marie Grant

Mathematics & Statistics Theses & Dissertations

Determination of molecular structure is commonly posed as a nonlinear optimization problem. The objective functions rely on a vast amount of structural data. As a result, the objective functions are most often nonconvex, nonsmooth, and possess many local minima. Furthermore, introduction of additional structural data into the objective function creates barriers in finding the global minimum, causes additional computational issues associated with evaluating the function, and makes physical constraint enforcement intractable. To combat the computational problems associated with standard nonlinear optimization formulations, Williams et al. (2001) proposed an atom-based optimization, referred to as GNOMAD, which complements a simple interatomic distance …

A Molecular Technique For Specific Identification Of Western Atlantic Ocean Scombrids And An Analysis Of A Larval Scombrid Assemblage Off The Kona Coast Of Hawaii, Melissa A. Paine

Dissertations, Theses, and Masters Projects

No abstract provided.

Altered Phosphorylation Of [Beta]-Catenin In Glucocorticoid Treated 235-1 Rat Pituitary Tumor Cells, Susie K. Saunders

Theses, Dissertations and Capstones

Beta-catenin is an essential cell adhesion and signaling protein, associated with high prolactin levels in rat pituitary tumor cells. It has been shown that phosphorylation affects the location and activity of b-catenin. Glycogen synthase kinase (GSK3-b) is a serine-threonine kinase that phosphorylates b-catenin on N-terminal residues, targeting it for proteasomal degradation. Studies have shown that C-terminal tyrosine phosphorylation decreases the association of b-catenin with cadherin. In 235-1 rat pituitary tumor cells, our lab has shown that the glucocorticoid analog dexamethasone (Dex) decreases the half- life of b-catenin while increasing the activity of GSK3-b. The current study was undertaken to examine …

A Mycobacterium-Inducible Nramp In Striped Bass Morone Saxatilis, Erin Jeffrey Burge

Dissertations, Theses, and Masters Projects

In mammals, the natural resistance-associated macrophage protein 1 gene, Nramp1, plays a major role in resistance to mycobacterial infections. Chesapeake Bay (USA) striped bass, Morone saxatilis, are currently experiencing an epizootic of mycobacteriosis that threatens the health of this ecologically and economically important species. This dissertation characterizes an Nramp gene in this species (MsNramp ) and provide evidence for induction following Mycobacterium exposure. The striped bass MsNramp gene and 554 amino acid sequence contain all the signal features of the Nramp family, including a topology of 12 transmembrane domains (TM), the transport protein specific 'binding-protein-dependent transport system inner membrane component …

Genetic Variation Within And Among The Suminoe Oyster (Crassostrea Ariakensis) Populations And Stocks As Assessed By Molecular Markers, Qian Zhang

Dissertations, Theses, and Masters Projects

No abstract provided.

Genetic Stock Structure Of The Sailfish, Istiophorus Platypterus, Based On Nuclear And Mitochondrial Dna, Jan Renee Mcdowell

Dissertations, Theses, and Masters Projects

The results of life history studies involving the sailfish, Istiophorus platypterus, from the Atlantic, Pacific, and Indian oceans were reviewed. Results of studies agree on several aspects of sailfish biology; sailfish exhibit opportunistic feeding behavior, follow the 28??C isotherm, spawn in summer mature at approximately age 3, and are the least migratory of the billfishes. However, results are in conflict concerning sex ratio and whether sailfish are single or multiple spawners. Although many researchers have reported that sailfish are sexually dimorphic, their fragile otoliths make them difficult to age, aging studies have not been validated, and sexual dimorphism has not …

Molecular Characterization Of Cultured Perkinsus Marinus Isolates, Gwynne D. Brown

Dissertations, Theses, and Masters Projects

Perkinsus marinus is the causative agent of the oyster disease Dermo in the eastern oyster, Crassostrea virginica. In vitro propagation of the parasite has led to the establishment of multiple isolates by several investigators. Little work, however, has been done to characterize different isolates. In this study multiple isolates were examined for genetic and biochemical diversity. at two loci, the ITS region and the ATAN region, there was as much intra-isolate genetic variation among DNA sequences of some isolates as there was inter-isolate variation. Variation was also observed at a third loci, a subtilisin-like serine protease gene. This is the …

Production Of Copper-Complexing Ligands In Response To Elevated Concentrations Of Copper In Marine Synechococcus Spp., Arunsri Chatkaeomorakot Brown

Theses and Dissertations in Biomedical Sciences

Synechococcus spp. (marine cyanobacteria) are extremely sensitive to copper toxicity and can produce high-affinity ligands of unknown structure which form complexes with free cupric ion. These ligands may contribute to the biological control of the levels of free cupric ions in surface seawater. Synechococcus spp. are known to produce metallothioneins (MT) in response to cadmium and zinc stress. In the present study the hypothesis that marine Synechococcus produce MT in response to copper was tested. Three marine Synechococcus spp. i.e., PCC 7001, PCC 73109, and PCC 7003, were exposed to different concentrations of CUSO 4 for various time periods. Size …

Environmental Stress In Hard Coral: Evaluating Lipid As An Indicator Of Sublethal Stress On Short Time Scales, David Harold Niebuhr

Dissertations, Theses, and Masters Projects

Lipid quality was evaluated in Montastrea spp. under sediment- and heat-stressed conditions to evaluate lipid ratio as an indicator of sub-lethal stress on short time scales. The ratio of storage lipid (wax ester + triacylglyceride) to structural lipid (sterol esters + phospholipid) decreased significantly (0.25 to 0.14, p < 0.01) after experimental sedimentation. FAME analysis of colonies exposed to experimental sedimentation showed a reduction of the algal, 18:3(n-6) and 18:4(n-3), polyunsaturated fatty acid (PUFA) in the polar lipid fraction. This loss of PUFA suggests a loss of algal membrane in sediment-stressed colonies. Lipid quality was similarly measured in M. faveolata over a 10-day period. Mean (n = 20) ratio of storage to structural lipid in M. faveolata dropped from 2.43 to a level of 0.98 immediately following a natural sedimentation event before recovering to levels of 1.4 and 2.9 on post-storm days 2 and 4, respectively. Colonies of M. annularis subject to heat-stress (35??C) exhibited no significant change in storage lipid ratio, while levels of Free Fatty Acids increased significantly from 0.012 (n = 22) to 0.156 mg lipid/g dry tissue (n = 22)(p < 0.05). FAME analysis of tissue lipids extracted from the heat-stressed colonies showed changes in the polar fraction, with significant decreases in the 18:3(n-6), 18:3(n-3), 18:4(n-3), 20:4(n-6) and 20:5(n-3) (p < 0.05) PUFA and subsequent significant increases in the saturated fatty acids, 16:0 and 18:0 (p < 0.05). These changes in lipid quantity and quality indicate possible oxidation and preferential digestion of zooxanthellar membranes. Stress experiments were repeated in M. annularis using VacutainerRTM blood collection tubes to collect micro-tissue samples without destroying skeleton of the sample colonies. A significant decrease in storage: structural lipid ratio after sedimentation was also detected using the micro-tissue technique. This study indicates that the relative abundance of lipid subclass components can indicate sub-lethal environmental stress, on short time scales, in M. annularis and M. faveolata. Furthermore, micro-tissue collection techniques permit repeated monitoring coral colonies to assess the manifestation of stress from first detection of impact at the cellular level to changes in community to changes in community structure detectable over longer time scales.