Biochemistry, Biophysics, and Structural Biology Commons^™

Effect Of Mutation Of Cvn7 Conservon Genes Sco6796 And Sco6798 Of Streptomyces Coelicolor., Caroline Fulmer

Undergraduate Distinction Papers

Abstract

Streptomyces coelicolor is a gram-positive Actinobacterium. It is a filamentous soil organism that performs complex multicellular development. Its physiological and morphological differentiation produces active secondary metabolite antibiotics that have pharmaceutical applications. Studies of S. coelicolor show that cyclic di-GMP proteins regulate physiological and morphological development. S. coelicolor has 13 copies of an operon that are collectively named “conservon” because they are highly conserved. The operons are named cvn1-13 and genes in each operon are denoted cvnA-D. RNA-Seq data of a cyclic di-GMP phosphodiesterase double mutant showed differential expression between wild type and many of the operons in the …

Identifying Rmda Protein Interactions In Streptomyces Using A Bacterial Two-Hybrid System, Rachel Nguyen

Undergraduate Distinction Papers

Streptomyces is a genus of the phylum actinobacteria most commonly found as soil bacteria and used as a major source of antibiotics. RmdA and RmdB are phosphodiesterases that break down the ubiquitous second messenger cyclic-di-GMP which determines colony morphology and development of Streptomyces. The objective of this research is to identify whether RmdA will have interactions with itself using the Bacterial Adenylate Cyclase Two-Hybrid (BACTH) System. Each gene was fused into one of two BACTH vectors that encode a different domain of a single protein (T18 and T25) and then cotransformed into the BACTH indicator strain. The transformants were …

The Identification And Characterization Of A Putative Chromosome Segregation Gene In Streptomyces Coelicolor, Sean Kirk

Undergraduate Distinction Papers

Streptomyces coelicolor is a soil bacterium that is a model for bacterial development. It is a filamentous, sporulating bacterium known to produce many medically utilized antibiotics. The goal of this research was to examine several developmental mutants and characterize novel genes of interest. Previously generated random transposon insertion mutants were analyzed using visual and microscopic phenotyping. Mutants of interest were further pursued and each transposon disruption site was identified by Inverse PCR and DNA sequencing. One of the novel genes is suspected to be involved in DNA segregation and codes for a putative membrane protein. Staining with propidium iodide was …

Shaping Light In Backward-Wave Nonlinear Hyperbolic Metamaterials, Thomas George, Alexander Popov, Sergey Myslivets, Vitaly Slabko, Victor Tkachenko

Chemistry & Biochemistry Faculty Works

Backward electromagnetic waves are extraordinary waves with contra-directed phase velocity and energy flux. Unusual properties of the coherent nonlinear optical coupling of the phase-matched ordinary and backward electromagnetic waves with contra-directed energy fluxes are described that enable greatly-enhanced frequency and propagation direction conversion, parametrical amplification, as well as control of shape of the light pulses. Extraordinary transient processes that emerge in such metamaterials in pulsed regimes are described. The results of the numerical simulation of particular plasmonic metamaterials with hyperbolic dispersion are presented, which prove the possibility to match phases of such coupled guided ordinary and backward electromagnetic waves. Particular …

Transmembrane Domains Of Highly Pathogenic Viral Fusion Proteins Exhibit Trimeric Association In Vitro, Stacy R. Webb, Stacy E. Smith, Michael G. Fried, Rebecca Ellis Dutch

Molecular and Cellular Biochemistry Faculty Publications

Enveloped viruses require viral fusion proteins to promote fusion of the viral envelope with a target cell membrane. To drive fusion, these proteins undergo large conformational changes that must occur at the right place and at the right time. Understanding the elements which control the stability of the prefusion state and the initiation of conformational changes is key to understanding the function of these important proteins. The construction of mutations in the fusion protein transmembrane domains (TMDs) or the replacement of these domains with lipid anchors has implicated the TMD in the fusion process. However, the structural and molecular details …

Phase Diagram Of Water Confined By Graphene, Zhenghan Gao, Nicolas Giovambattista, Ozgur Sahin

Publications and Research

The behavior of water confined at the nanoscale plays a fundamental role in biological processes and technological applications, including protein folding, translocation of water across membranes, and filtration and desalination. Remarkably, nanoscale confinement drastically alters the properties of water. Using molecular dynamics simulations, we determine the phase diagram of water confined by graphene sheets in slab geometry, at T = 300 K and for a wide range of pressures. We find that, depending on the confining dimension D and density σ, water can exist in liquid and vapor phases, or crystallize into monolayer and bilayer square ices, as observed in …

Cation Exchange Capacity Determination: Comparative Study Of Three Methods, Lillie Zech

Undergraduate Research Conference

No abstract provided.

Assay Of The Reverse Osmosis Purified Water In The Life Science Building At Bowling Green State University, Ohio, Nicholas Mendenhall, Scott O. Rogers, Neocles B. Leontis

Honors Projects

Contaminated water sources can cause problems for scientific research and result in costly delays and failures of experiments. At Bowling Green State University, the reverse osmosis supply circulating in the Life Sciences Building has been measurably contaminated for nearly three years, corresponding to a change in servicing of the system. While servicing has been accelerated, the contamination in the system remains. The focus of this research was to identify the species of bacteria and fungi growing inside of the water system so that it might alert those servicing the system, and to begin to eliminate the contamination. Reverse osmosis water …

Data-Driven Computational Approach To Study Bio-Molecular Interactions, Misagh Naderi

LSU Doctoral Dissertations

Proteins commonly convey their functions in coordination with other proteins, small molecules and/or other biological assemblies such as sugars, lipids, DNA or RNA. Understanding the nature of these interactions is therefore central to improving our knowledge of biological systems. This body of work is a consolidation of three different computational approaches to study bio-molecular interactions: large-scale protein-ligand modeling, fragment-based cheminformatics, and computational analysis of interactions in single proteins.

The first issue that is addressed in this study is the scarcity of atomic crystal structures of protein-drug complexes. In general, a drug molecule’s affinity for multiple protein targets may causes unsolicited …

Synthesis Of Oxygenated Boronic Acid Substituted A-Cyanostilbenes For Use As Antibacterial, Yewon Kim

Honors Theses

Stilbenes are naturally occurring compounds that have exhibited antibacterial activity, although the biological effect of various stilbenes differs for Gram-positive and Gram-negative strains of bacteria. In prior research, cyanostilbenes have shown slight antibacterial activity (Brownlee, et al. 1943). This project aims to explore whether hybrid oxygenated a-cyanostilbenes possessing a boronic acid pharmacophore exhibits significant antibacterial activity (Das et al. 2013). The biological activity against Staphylocococcus aureus (Gram-positive) and Escherichia coli (Gram-negative) was tested using the Kirby-Bauer text. No inhibition of E. coli growth was shown while S. aureus was partially inhibited.

Evaluating Mutagenicity Of Burned Arginine-Based Heterocyclic Amines And Anti-Mutagenicity Effect Of Chinese Medicinal Herbs, Rayford C. Alva

Honors Theses

From the unexpected finding that cooked grains and meat substitutes elicit a mutagenic response in Salmonella typhimurium TA98, our work has been aimed at deconstructing this finding via a survey of heated binary amino acid combinations, involving arginine, a plant-based amino acid. secondly, our work has looked towards sing phytochemical extracts from traditional Chinese medicinal herbs to inhibit the mutagenic activity of heterocyclic amines (main culprit for mutagenicity in meat). Two fractions from the burned products of arginine and phenylalanine (RF-HCA-06 and -07) were statistically significant inducers of mutagenesis; Scutellaria barbata and Oldenlandia diffusa, both separately and together, were statistically …

Investigation Of Alcohol-Induced Changes In Hepatic Histone Modifications Using Mass Spectrometry Based Proteomics, Crystina Leah Kriss

USF Tampa Graduate Theses and Dissertations

Alcohol liver disease (ALD) is a major health concern throughout the world. Currently, in the United States, 17 million people suffer from alcoholism, of which 1.4 million people are receiving treatment [1, 2]. The link between ethanol metabolism, reactive oxygen species (ROS) and liver injury in ALD has been well characterized over the last couple decades [3-10]. Ethanol metabolism relies on the availability of the cofactor NAD+ for the oxidation of ethanol into acetate, consequently causing alterations in redox potential. Redox dysfunction within the mitochondria can affect multiple pathways important in maintaining cellular homeostasis. Chapter 1 provides an introduction to …

The Role Of Phosphohistidine Phosphatase 1 In Ethanol-Induced Liver Injury, Daniel Richard Martin

USF Tampa Graduate Theses and Dissertations

Chronic liver diseases, which includes alcoholic liver disease (ALD), are consistently among the top 15 leading causes of death in the United States. ALD is characterized by progression from a normal liver to fatty liver disease (hepatic steatosis), which can lead to cirrhosis, alcoholic hepatitis, and liver failure. We have identified a novel role of phosphohistidine signaling, mediated through phosphohistidine phosphatase 1 (PHPT1), in the onset of hepatic steatosis. We have identified PHPT1 as a target of selective oxidation following acute ethanol exposure as well as being downregulated following chronic ethanol exposure. We mapped the oxidative modification site and developed …

Elemental Analysis Of Beer Manufactured In Virginia Satellite Breweries, Jennifer Wiliams

Student Scholar Showcase

Financial incentives from the state and wider distribution potential have lured many west coast brewers to open satellite breweries in Virginia. Quality control is crucial for any brewery and especially important in satellite breweries that need to imitate the standard set by the parent brewery. Zinc, magnesium, iron, sodium manganese, and calcium are trace elements found in wort and are essential for enzymes that catalyze fermentation reactions. These elements protect against cell stress, play a crucial role in flocculation, and can affect the flavoring and coloration of beer. Barley, water, and hops are the sources of these elements in beer. …

Whole Tart Cherry Improves Disease Activity Index In Rat Model Of Dextran Sulfate Solution – Induced Ulcerative Colitis By Downregulating The Janus 1 And Janus 3 Kinases And Tnf - Alpha, Johana Alexandra Coronel

LSU Master's Theses

Ulcerative colitis (UC) is a disabling inflammatory bowel disease. UC is characterized by chronic, relapsing inflammation of the colon and rectum. Current treatments such as monoclonal antibodies against TNF-α, IL-6, IL-12/p40, adhesion molecules, Janus kinases (JAK) inhibitors have side effects or lose their effects over time. Alternative approaches with fewer side effects for patients are needed. Montmorency tart cherries (Prunus cerasus) are a good source of anti-inflammatory flavonoids. We hypothesized that regular consumption whole tart cherry (TC) standardized to its major anthocyanin content, namely cyanidin-3-glucosyl-rutinoside, would be effective in reducing inflammation in UC. The aim of this research was to …

Cloning, Purification, And Preliminary Dna-Binding And Unfolding Results For The Dna Polymerase I From The Psychrophile Psychromonas Ingrahamii, John Tod Baker

LSU Master's Theses

Psychromonas ingrahamii is a psychrophilic bacterium that lives in Arctic polar sea ice and grows at a temperature range of -12 to 10º C. This bacterium resides within veins inside the ice where the salinity is high, resulting in a freezing point depression and liquid water. The large fragment of DNA polymerase I from Psychromonas ingrahamii, called Klenpin, has been cloned, expressed, and purified in our laboratory. Although enzyme kinetic studies have been performed on a few psychrophilic enzymes, the thermodynamics of ligand binding and of protein stability have not been well studied for this class of extremophilic proteins. …

Analyzing The Effects Of Ca2+ Dynamics On Mitochondrial Function In Health And Disease, Patrick Toglia

USF Tampa Graduate Theses and Dissertations

Mitochondria plays a crucial role in cells by maintaining energy metabolism and directing cell death mechanisms by buffering calcium (Ca²⁺ )from cytosol. Therefore, the Ca²⁺ overload of mitochondria due to the upregulated cytosolic Ca²⁺ , observed in many neurological disorders is hypothesized to be a key pathway leading to mitochondrial dysfunction and cell death. In particular, Ca²⁺ homeostasis disruptions due to Alzheimer’ s disease (AD)-causing presenilins (PS1/PS2) and oligomeric forms of β-amyloid peptides Aβ commonly found in AD patients are presumed to cause detrimental effects on the mitochondria and its ability to function properly. We begin …

Elastin-Like Polypeptide Fusion Tag As A Protein-Dependent Solubility Enhancer Of Cysteine-Knot Growth Factors, Tamina L. Johnson

USF Tampa Graduate Theses and Dissertations

Elastin-like peptide (ELP) fusions promote therapeutic delivery and efficacy. Recombinant proteins, like neurotrophins, lack bioavailability, have short in vivo half-lives, and require high manufacturing costs. Fusing recombinant proteins with genetically encodable ELPs will increase bioavailability, enhance in vivo solubilization, as well as provide a cost-effective method for purification without the need for chromatography. During expression of neurotrophin-ELP (N-ELP) fusions, dense water-insoluble aggregates known as inclusion bodies (IBs) are formed. Inclusion bodies are partially and misfolded proteins that usually require denaturants like Urea for solubilization. Strong denaturants arrest ELPs stimuli-responsive property and increase unwanted aggregation, making purification difficult, yet possible. The …

Anti-Fungal Compounds From Native Prairie Plants, Sarah Mcmichael, Kirk Manfredi

Research in the Capitol

Plant compounds can be found in many consumer products such as nutritional supplements (nutraceuticals), soaps, shampoos, and other personal care products. They are not yet used in these products as preservatives. This area of research focusses on identifying compounds from native prairie plants that can be used as preservatives in personal care products. An objective of this research is to isolate compounds that can be used to replace synthetic parabens. Synthetic parabens have been found to be harmful to humans and the environment. Our isolate would be a natural preservative and eliminate the harmful synthetic. This research presentation will discuss …

Ef-P Post-Translational Modification Has Variable Impact On Polyproline Translation In Bacillus Subtilis, Anne Witzky, Katherine R. Hummels, Rodney Tollerson Ii, Andrei Rajkovic, Lisa A. Jones, Daniel B. Kearns, Michael Ibba

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Elongation factor P (EF-P) is a ubiquitous translation factor that facilitates translation of polyproline motifs. In order to perform this function, EF-P generally requires posttranslational modification (PTM) on a conserved residue. Although the position of the modification is highly conserved, the structure can vary widely between organisms. In Bacillus subtilis, EF-P is modified at Lys³² with a 5-aminopentanol moiety. Here, we use a forward genetic screen to identify genes involved in 5-aminopentanolylation. Tandem mass spectrometry analysis of the PTM mutant strains indicated that ynbB, gsaB, and ymfI are required for modification and that yaaO, yfkA, and …

Evaluation Of Controlled-Release Nitrogen And Potassium Fertilizer In Louisiana Sugarcane Production, Samuel Kwakye

LSU Master's Theses

Sugarcane (Saccharum spp) production is an integral part of Louisiana's economy. Proper fertilizer management, particularly nitrogen (N) and potassium (K), optimizes sugarcane production. An experiment was established in 2015 at the LSU AgCenter Sugar Research Station in St. Gabriel, Louisiana to: (1) assess the effect of N and K fertilizer sources on sugarcane yield and quality parameters, and (2) monitor the changes in inorganic N and K levels in the soil at different sugarcane growth stages. The experiment was arranged in a randomized complete block design on a Sharkey clay soil and complete randomized design on a Commerce …

Breaking Conventions: Ohrr, A Transcriptional Regulator From Burkholderia Thailandensis, Anuja Rajiv Pande

LSU Doctoral Dissertations

Organic peroxides (OPs) are highly toxic oxidants. They directly react with cellular macromolecules rendering them inactive. Soil bacteria live in an oxidative environment rich in OPs from plant exudates and other xenobiotic compounds. Bacteria also face a burst of organic peroxides (lipid peroxides) in the macrophages during host invasion. Prokaryotes sense oxidative changes in environment and modulate their gene expression as an adaptive response for survival. MarR (multiple antibiotic resistance regulators) proteins enable bacteria to respond to such stress. Resistance to organic oxidants has been hypothesized to help bacteria survive in presence of disinfectants. This increases nosocomial infection rate.

Bacteria …

Vesicles Mimicking Normal And Cancer Cell Membranes Exhibit Differential Responses To The Cell-Penetrating Peptide Pep-1, Bashiyar Almarwani, Esther Nzuzi Phambu, Christopher Alexander, Ha Aimee T. Nguyen, Nsoki Phambu, Anderson Sunda-Meya

Chemistry Faculty Research

The cell-penetrating peptide (CPP) Pep-1 presents a great potential in drug delivery due to its intrinsic property to cross plasma membrane. However, its mechanism of entry into the cell remains unresolved. In this study, we compare the selectivity of Pep-1 towards vesicles mimicking normal and cancer cell membranes. The interaction was performed in a wide range of peptide-to-lipid molar ratios using infrared (IR), fluorescence, scanning electron microscopy (SEM), thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) techniques. At low peptide concentration, fluorescence experiments show that lipid-phosphatidylserine (PS) seems to enable Pep-1 translocation into cancer cell membrane as evidenced by the …

Changes In Carbon, Nutrient, And Trace Metal Concentrations During The Transition From Freshwater To Marine Environments - A Case Study In Southwest Louisiana, Songjie He

LSU Doctoral Dissertations

Rivers carry terrestrial nutrients and other materials to estuaries, affecting biogeochemical cycles and ecosystems in coastal margins. This dissertation research incorporated five studies to investigate the dynamics of carbon, nutrients, and trace metals in four major coastal rivers in southwest Louisiana – the Sabine, Calcasieu, Mermentau, and Vermilion Rivers. The overarching goal of this research was to broaden the knowledge and understanding of biogeochemical processes in freshwater – saltwater mixing zones. These studies employed decade-long monitoring records of water chemistry, utilized monthly field measurements conducted over a two-year period, and used analysis of over 200 water samples for carbon, nitrogen, …

Biomimetic Studies Of Oxidation Reactions By Metalloporphyrins Through Ligand Effect And Kinetic Studies Of Photo-Generated Porphyrin-Iron(Iv)- Oxocompound Ii Models, Dharmesh J. Patel

Masters Theses & Specialist Projects

High-valent iron(IV)-oxo porphyrins are the central oxidizing species in hemecontaining enzymes and synthetic oxidation catalysts. Many transition metal complexes have been extensively studied as models of the ubiquitous cytochrome P450 enzymes to probe the sophisticated oxygen atom transfer (OAT) mechanism as well as to invent enzyme-like oxidation catalysts. In this work, two metalloporphyrin complexes have been successfully synthesized, and spectroscopically characterized. A new photochemical entry to porphyrin-iron(IV)-oxo derivatives, commonly referred to as compound II models, was also investigated in two porphyrin ligands that differ in electronic and steric environments. As determined by their distinct UV-vis spectra and kinetic behaviors, iron(IV)-oxo …

Untargeted Bioassay Strategy For Medicinal Plants: In Vitro Antidiabetic Activity And 13c Nmr Profiling Of Extracts From Vitex Negundo L, Fabian M. Dayrit, Myrnille Joy B. Zabala, Lolita G. Lagurin

Chemistry Faculty Publications

Bioassay-guided fractionation is the principal method for the identification of active constituents in medicinal plants. By design, this method aims to identify the most active compound in a complex mixture with the objective of discovering novel drug candidates. Described here is a complementary method for the identification of known bioactive compounds in medicinal plants which is untargeted and which takes advantage of the large NMR database of known natural products and availability of statistical software. This untargeted bioassay strategy is demonstrated as a proof of principle in the determination of the antidiabetic compounds in Vitex negundo L. Crude methanol and …

Toxicological Assessment Of Nanomaterials: The Role Of In Vitro Raman Microspectroscopic Analysis, Esen Efeoglu, Marcus Maher, Alan Casey, Hugh Byrne

Articles

The acceleration of nanomaterials research has brought about increased demands for rapid analysis of their bioactivity, in a multi-parametric fashion, to minimise the gap between potential applications and knowledge of their toxicological properties. The potential of Raman microspectroscopy for the analysis of biological systems with the aid of multivariate analysis techniques has been demonstrated. In this study, an overview of recent efforts towards establishing a ‘label-free high content nanotoxicological assessment technique’ using Raman microspectroscopy is presented. The current state of the art for cellular toxicity assessment and the potential of Raman microspectroscopy are discussed, and the spectral markers of the …

Magnetic Properties Of Acenes And Their O-Quinone Derivatives: Computer Simulation, A. A. Starikova, A. G. Starikov, R. M. Minyaev, Alexander I. Boldyrev, V. I. Minkin

Chemistry and Biochemistry Faculty Publications

Quantum chemical study (DFT UB3LYP/6-311++G(d,p)) of the structure and properties of acenes functionalized with two o-benzoquinone groups and their complexes with sodium cations has been performed. An increase in the number of fused rings has been shown to result in the stabilization of biradicaloid state of acenes and the switching of the character of exchange interactions between redox-active moieties from antiferromagnetic to ferromagnetic. The obtained results allow one to consider o-quinone acene derivatives as a basis for designing magnetoactive compounds.

Monitoring And Quantifying Tetracycline Resistance Genes In A Swine Waste Anaerobic Digester Over A 100-Day Period, Melanie Couch

Masters Theses & Specialist Projects

Unregulated use of growth promoting antibiotics like Tetracyclines in agricultural feeds is becoming an increasing problem in antibiotic resistance. Undigested antibiotics leads to significant concentrations in livestock waste. These concentrations provide continuous selection pressure for the development of antibiotic resistance genes in the environment. Antibiotic resistance related deaths are projected to surpass cancer related deaths by 2050 making antibiotic resistance a pressing public health issue. The purpose of this study is to determine the abundance and persistence of tetracycline (tet) resistance genes in swine waste over a period of 100 days in an anaerobic digester system. Tet(A), tet(B), tet(G), tet(M), …

Detection Of Amyloid Beta (Aβ) Oligomeric Composition Using Matrix-Assisted Laser Desorption Ionization Mass Spectrometry (Maldi Ms)., Jasmine S-H Wang, Shawn N Whitehead, Ken K-C Yeung

Chemistry Publications

The use of MALDI MS as a fast and direct method to detect the Aβ oligomers of different masses is examined in this paper. Experimental results suggest that Aβ oligomers are ionized and detected as singly charged ions, and thus, the resulting mass spectrum directly reports the oligomer size distribution. Validation experiments were performed to verify the MS data against artifacts. Mass spectra collected from modified Aβ peptides with different propensities for aggregation were compared. Generally, the relative intensities of multimers were higher from samples where oligomerization was expected to be more favorable, and vice versa. MALDI MS was also …