Biochemistry Commons^™

Remote Performance Modulation Of Ultrafiltration Membranes By Magnetically And Thermally Responsive Polymer Chains, Arijit Sengupta, Anh Vu, Xianghong Qian, Sumith R. Wickramasinghe

Chemical Engineering Faculty Publications and Presentations

Ultrafiltration membranes, that respond to an external magnetic field and local temperature have been developed. Surface-initiated activator-generated electron transfer (AGET) atom transfer radical polymerization (ATRP) has been used to graft poly(N-isopropylacrylamide) (PNIPAm) from the surface of 300 kDa regenerated cellulose membranes. The polymerization initiator was selectively attached to the entire membrane surface, only the outer membrane surface or only the inner pore surface. A superparamagnetic nanoparticle was attached to the end of the polymer chain. The DI water flux as well as the flux and rejection of bovine serum albumin were investigated in the absence and presence of a 20 …

The Discovery And Analysis Of Peroxidase Enzyme In Pueraria Montana, Kathryn Briley

Senior Honors Theses

Peroxidase enzymes are used for a variety of industrial and biotechnological applications because of their ease of purification, broad range of chemical activities, and low cost of use. Identification of quality peroxidase sources that are convenient for enzymatic isolation and give way to high yields of product is a desirable pursuit in biochemical research. Kudzu is an excellent candidate for this pursuit as it displays high catalytic activity in screening assays and is found in abundance. This project seeks to determine the enzyme stability, optimal pH conditions, and possible novel chemical activities of peroxidase isolated from kudzu leaves. The methods …

An Investigation Of K2 Mycobacteriophage Lysin A Proteins, Ethan Dionne

Chemistry & Biochemistry Student Scholarship

Major: Biochemistry
Faculty Mentor: Dr. Kathleen Cornely, Chemistry and Biochemistry

Investigations Into The Cellular Target Of 4-Trifluoromethoxy Chalcone Via Darts Method, Jordan Stacy

Undergraduate Theses

Cellular drug target discovery is an important step in any drugs journey from bench to bedside. This is true for our lab's molecule of interest, the Chalcone. The Chalcone molecule and its derivatives have been identified as small, plant-derived secondary metabolites that, when interacting with human cancer cell lines, trigger apoptotic pathways leading to varying levels of cell death. One derivative, 4-Trifluoromethoxy Chalcone (4TFM), was identified through screenings as inducing the highest death rate in A549 cancer cells, in conjunction with having the lowest IC50, making it a good candidate to use in searching for the currently unknown cellular target …

Antibacterial And Antioxidant Potential Of A Novel Plant, Eriodictyon Californicum, And Its Healing Abilities, Allie Richards

Undergraduate Theses

Our body is under constant attack of oxidative stress (OS). OS is a crucial factor in the pathogenesis and progression of various chronic diseases such as diabetics, rheumatoid arthritis, myocardial infarction, cardiovascular diseases, stroke, cancer, Alzheimer’s disease, Parkinson’s disease, aging and more. The main cause of OS is free radicals, which are continuously generated in our body during normal metabolic processes. These free radicals have the potential to damage any of our key biomolecules. To protect ourselves from the toxic effect of free radicals the human body has a built-in army of antioxidants that play an important role in the …

Europium Sulfide Nanoprobes Predict Antiretroviral Drug Delivery Into Hiv-1 Cell And Tissue Reservoirs, Jonathan Herskovitz, Mahmudul Hasan, Jatin Machhi, Insiya Mukadam, Brendan M. Ottemann, James R. Hilaire, Christopher Woldstad, Joellyn Mcmillan, Yutong Liu, Javier Seravalli, Anandakumar Sarella, Howard E. Gendelman, Bhavesh D. Kevadiya

Department of Biochemistry: Faculty Publications

Background: Delivery of long-acting nanoformulated antiretroviral drugs (ARVs) to human immunodeficiency virus type one cell and tissue reservoirs underlies next generation antiretroviral therapeutics. Nanotheranostics, comprised of trackable nanoparticle adjuncts, can facilitate ARV delivery through real-time drug tracking made possible through bioimaging platforms. Methods: To model HIV-1 therapeutic delivery, europium sulfide (EuS) nanoprobes were developed, characterized and then deployed to cells, tissues, and rodents. Tests were performed with nanoformulated rilpivirine (NRPV), a non-nucleoside reverse transcriptase inhibitor (NNRTI) used clinically to suppress or prevent HIV-1 infection. First, CD4+ T cells and monocyte-derived macrophages were EuS-treated with and without endocytic blockers to identify …

The Analysis Of Folate-Dependent Transcription Factor Zinc Finger Protein 410, Feifan Xu

Senior Honors Theses

A previous study that introduced dietary folate to mice in the form of folic acid to determine if gene activity would be altered based on this biological molecule demonstrated that mice without folic acid had cognition deficits, and this phenomenon was correlated with altered gene expression in their brains. The included bioinformatic analysis revealed two main transcription factors that bind to proteins in the nucleus, and one is known as the Zinc Finger Protein 410 (Zfp410). Due to the lack of literature explaining the function of this transcription factor, this project is intended to analyze Zfp410 in detail from scratch. …

216— Loss Of Function Mutation For Tp53 Does Not Rescue The Chaf1bNt2 Small-Eye Phenotype In Danio Rerio, Alex Parks

GREAT Day Posters

In Zebrafish, the chromosome assembly factor 1b (chaf1b) gene is in part responsible for the development of the eye. In homozygous chaf1bt24412 mutants retinal cell death is promoted through cell-death promoting activity of the gene, tumor suppressor protein p53 (tp53), resulting in a small-eye phenotype. Another allele chaf1bnt2, was found to also result in the small-eye phenotype when in a homozygous state. We found that knockdown of Tp53 protein via morpholino antisense oligonucleotide injection of 1-2 cell stage embryos failed to rescue retinal cell death of chaf1bnt2 homozygous mutants as detected by TUNEL labeling. Because morpholinos may fail to fully …

Using The Lambert Function To Map Enzyme Kinetics, Melissa Gouge

ONU Student Research Colloquium

In an academic setting, enzyme kinetics are introduced by analysis of the initial rates using the Michalis-Menten model. To better understand enzyme kinetics, a complete time course analysis will provide a larger set of information. The use of the integrated form of the rate equation allows for a beginning glance at the mechanism of reaction. With this, the Lambert Ω function can be used to analyze the integrated rate equations. There are two different branches of the Lambert Ω function, a negative branch and a positive branch. For each branch there is a unique expanded series approximation used for analysis …

The Effect Of Cadmium On Ovarian Adenocarcinoma Cell Lines: An Investigation Of The Possible Mechanism Of Action, Haley Todd

Undergraduate Theses

Cadmium, a heavy metal and carcinogen, is an environmental and workplace contaminate. As a known endocrine disruptor, it can mimic the proliferative effects of estrogen and is classified as a metalloestrogen. While the proliferative effect of estrogen on cancerous cell growth has been well established, the effects of cadmium have not been fully examined. To determine if cadmium stimulates growth in two human ovarian adenocarcinoma cell lines, OVCAR3 and SKOV3, cells were treated for 48 hours with varying concentrations of cadmium, 0.001 µM – 10 µM, and growth was measured using a cell proliferation assay. Both cell lines showed a …

Crispr/Cas9-Induced Fad2 And Rod1 Mutations Stacked With Fae1 Confer High Oleic Acid Seed Oil In Pennycress (Thlaspi Arvense L.), Brice A. Jarvis, Trevor B. Romsdahl, Michaela G. Mcginn, Tara J. Nazarenus, Edgar B. Cahoon, Kent D. Chapman, John C. Sedbrook

Department of Biochemistry: Faculty Publications

Pennycress (Thlaspi arvense L.) is being domesticated as an oilseed cash cover crop to be grown in the off-season throughout temperate regions of the world. With its diploid genome and ease of directed mutagenesis using molecular approaches, pennycress seed oil composition can be rapidly tailored for a plethora of food, feed, oleochemical and fuel uses. Here, we utilized Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology to produce knockout mutations in the FATTY ACID DESATURASE2 (FAD2) and REDUCED OLEATE DESATURATION1 (ROD1) genes to increase oleic acid content. High oleic acid (18:1) oil is valued for its oxidative stability that is …

Funtionalization Of Morphologically Different Nanoclays For Enhanced Mechanical Properties, Malachi Kent

Honors College Theses

Nanoclays are microscopic clays that are comprised of layered nanoparticle aluminosilicates that are naturally arranged in several morphologies. Halloysite nanotubes (HNT) are hollow, double-layered tubes, bentonite (BNT) clays are trapezoidal platelets, and nanosilica or silica dioxide (SiO₂) are spherical. These nanoclays were chosen for sustainable purposes as they are naturally derived and for economical purposes because of their cheap commercial availability. In this study, the nanoclays are functionalized with an epoxy organosilane group through reflux conditions at 120 °C for 24 hours. The pristine and functionalized nanoclays were extensively characterized with infrared radiation (IR) spectroscopy, scanning electron microscope …

Studies Of The Rectification Behavior Of The Pdt Ligand And The Active Site Of Msrp, Laura J. Ingersol

Chemistry and Chemical Biology ETDs

Molybdenum (Mo) is an essential element that plays an important role in global nitrogen, carbon, and sulfur cycles with a critical role in human metabolism and ecological balance. It becomes catalytically active when complexed with the pyranopterin dithiolene ligand (PDT), forming the nearly ubiquitous molybdenum cofactor (Moco). The complex biosynthetic pathway of Moco, its presence in all three domains of life, and its role as a constituent cofactor in the last universal common ancestor (LUCA) all point toward the importance of the PDT in the development of life on Earth. Molybdoenzymes catalyze the two-electron oxidation or reduction of substrates that …

Project Alien, Rebecca Mccallin, Madelyn Hoying, Alex Evans, Matthew Nestler, Karli Rae Sutton, Garett Craig, Lucia Secaia Del Cid, Alexander Guy, Rachel Fernandez, Amanda Trusiak, Paige Aley, Ingabire Gakwerere, Nina Dorfner, Maria Mosbacher, Mary Flavin, Selvin Hernandez, Audrey Steen, Benjamin Kazimer

Undergraduate Research and Scholarship Symposium

Project ALIEN is a comprehensive plan to send humans to Mars to look for life on the Martian surface while exploring the viability and adaptability of terrestrial microbes in Martian atmospheric conditions. ALIEN will use a ballistic capture trajectory to get to Mars and stay in aerostationary orbit for a 30-day surface mission, during which two surface crewmembers will perform a variety of experiments to achieve the mission’s goals of Martian microbial discovery within brines of the Gale Crater and terrestrial microbe adaptability and viability to Martian conditions. Experimentation is based on the presumption that Martian microbes are metabolically similar …

Bacteria, Guano And Soot: Source Assessment Of Organic Matter Preserved In Black Laminae In Stalagmites From Caves Of The Sierra De Atapuerca (N Spain), Joeri Kaal, Virginia Martínez-Pillado, Antonio Martínez Cortizas, Jorge Sanjurjo Sánchez, Arantza Aranburu, Juan-Luis Arsuaga, Eneko Iriarte

International Journal of Speleology

Speleothems are a recognized source of paleoclimatic information, but their value as a source of signals from human activities in caves with an archaeological record has rarely been explored. Previous studies of speleothems in the Sierra de Atapuerca karst system (Burgos, northern Spain) revealed an important human fossil record, provided information about human activities in and around these caves, and the impacts on their natural environment. The present study reports the results of molecular characterization of dark-colored laminae from the stalagmites Ilargi (Galería de las Estatuas) and GS1, GS2, and GS3 (Galería del Silo), by pyrolysis-GC-MS (Py-GC-MS) and …

The Supercam Instrument Suite On The Mars 2020 Rover: Science Objectives And Mast-Unit Description, S. Maurice, R. C. Wiens, P. Bernardi, P. Caïs, S. Robinson, T. Nelson, O. Gasnault, J. -M. Reess, M. Deleuze, F. Rull, J. -A. Manrique, S. Abbaki, R. B. Anderson, Y. André, Stanley M. Angel, G. Arana, T. Battault, P. Beck, K. Benzerara, S. Bernard, Et. Al.

Faculty Publications

On the NASA 2020 rover mission to Jezero crater, the remote determination of the texture, mineralogy and chemistry of rocks is essential to quickly and thoroughly characterize an area and to optimize the selection of samples for return to Earth. As part of the Perseverance payload, SuperCam is a suite of five techniques that provide critical and complementary observations via Laser-Induced Breakdown Spectroscopy (LIBS), Time-Resolved Raman and Luminescence (TRR/L), visible and near-infrared spectroscopy (VISIR), high-resolution color imaging (RMI), and acoustic recording (MIC). SuperCam operates at remote distances, primarily 2–7 m, while providing data at sub-mm to mm scales. We report …

Broken-Hearted” Carbon Bowl Via Electron Shuttle Reaction: Energetics And Electron Coupling, Gabrielle A. Leith, Allison M. Rice, Brandon J. Yarbrough, Preecha Kittikhunnatham, Abijai Mathur, Nicholas A. Morris, Megan J. Francis, Anna A. Berseneva, Poonam Dhull, Richard D. Adams, M. Victoria Bobo, Aaron A. Vannucci, Mark D. Smith, Sophya Garashchuk, Natalia B. Shustova

Faculty Publications

Unprecedented one-step CC bond cleavage leading to opening of the buckybowl (π-bowl), that could provide access to carbon-rich structures with previously inaccessible topologies, is reported; highlighting the possibility to implement drastically different synthetic routes to π-bowls in contrast to conventional ones applied for polycyclic aromatic hydrocarbons. Through theoretical modeling, we evaluated the mechanistic pathways feasible for π-bowl planarization and factors that could affect such a transformation including strain and released energies. Through employment of Marcus theory, optical spectroscopy, and crystallographic analysis, we estimated the possibility of charge transfer and electron coupling between “open” corannulene and a strong electron acceptor such …

Isoprene Production From Municipal Wastewater Biosolids By Engineered Archaeon Methanosarcina Acetivorans, Sean Carr, Jared Aldridge, Nicole R. Buan

Department of Biochemistry: Faculty Publications

Wastewater biosolids are a promising feedstock for production of value-added renewable chemicals. Methane-producing archaea (methanogens) are already used to produce renewable biogas via the anaerobic treatment of wastewater. The ability of methanogens to efficiently convert dissolved organic carbon into methane makes them an appealing potential platform for biorefining using metabolic engineering. We have engineered a strain of the methanogen Methanosarcina acetivorans to produce the volatile hemiterpene isoprene in addition to methane. The engineered strain was adapted to grow in municipal wastewater through cultivation in a synthetic wastewater medium. When introduced to municipal wastewater the engineered methanogens were able to compete …

Isoprene Production From Municipal Wastewater Biosolids By Engineered Archaeon Methanosarcina Acetivorans, Sean Carr, Jared Aldridge, Nicole R. Buan

Department of Biochemistry: Faculty Publications

Wastewater biosolids are a promising feedstock for production of value-added renewable chemicals. Methane-producing archaea (methanogens) are already used to produce renewable biogas via the anaerobic treatment of wastewater. The ability of methanogens to efficiently convert dissolved organic carbon into methane makes them an appealing potential platform for biorefining using metabolic engineering. We have engineered a strain of the methanogen Methanosarcina acetivorans to produce the volatile hemiterpene isoprene in addition to methane. The engineered strain was adapted to grow in municipal wastewater through cultivation in a synthetic wastewater medium. When introduced to municipal wastewater the engineered methanogens were able to compete …

Coilin Expression During Nucleolar Stress In The Drosophila Brain, Kathryn Rose Deleo

LSU Master's Theses

Impairment of ribosome biogenesis disrupts cellular homeostasis and, in the process, causes a state of nucleolar stress in a cell. This triggers human syndromes collectively known as ribosomopathies. One such ribosomopathy is the Treacher Collins syndrome (TCS) which causes congenital craniofacial birth defects and hearing loss in patients. In TCS migration of neural crest cells during development is hindered by the loss of the ribosome assembly factor, treacle, and consequent apoptosis of these progenitor cells. Modeling this disorder in Drosophila was previously accomplished by disruption of Nopp140, treacle’s closest equivalent in Drosophila. During nucleolar stress, the Mushroom Body neuroblast …

Structural Basis For Clostridium Perfringens Enterotoxin Targeting Of Claudins At Tight Junctions In Mammalian Gut, Alex J. Vecchio, Sewwandi S. Rathnayake, Robert M. Stroud

Department of Biochemistry: Faculty Publications

The bacterium Clostridium perfringens causes severe, sometimes lethal gastrointestinal disorders in humans, including enteritis and enterotoxemia. Type F strains produce an enterotoxin (CpE) that causes the third most common foodborne illness in the United States. CpE induces gut breakdown by disrupting barriers at cell–cell contacts called tight junctions (TJs), which are formed and maintained by claudins. Targeted binding of CpE to specific claudins, encoded by its C-terminal domain (cCpE), loosens TJ barriers to trigger molecular leaks between cells. Cytotoxicity results from claudin-bound CpE complexes forming pores in cell membranes. In mammalian tissues, ∼24 claudins govern TJ barriers—but the basis for …

Thermodynamic Analysis Of Pamam Dendrimer Synthesis, Ryutaro Einar Jacobson

Honors Theses

The objective of this research was to determine kinetic and thermodynamic data about the reaction between ethylene diamine and methyl acrylate which is used in the production of PAMAM dendrimers. Establishing a rate law and finding values such as rate constant, activation energy, and Arrhenius coefficient will help in determining the best possible conditions for the reaction. A rate law was established through the method of initial rates, comparing the rates of reactions with different initial concentrations to one another. Additional reactions were run with active cooling to observe the effects on rate and reaction progress, and finally reactions were …

Finding Optimal Opa-Iblc Derivatized Amino Acid Analysis Parameters With Three-Dimensional Fluorometric Scans For Hplc Method Validation, Aaron Jacobs

Honors Theses

Three-dimensional fluorometric scans of OPA-IBLC derivatized glutamic acid and histidine solutions revealed that decreasing concentration maintained maximum λem but produced different maximum λex. In pH 0.4 M sodium borate buffer (pH 10.4), 50 μM derivatized amino acid solutions had maxima around 350- to 450-nm λex-λem where 12.5 μM solutions had maxima around 230- to 450-nm λex- λem. In the lower concentration, the λex peak at 230 nm was about 80% higher than 350 nm. Thus, collecting spectra with 230 nm λex would provide better LOD and LOQ. Results inform HPLC standard concentrations since differential quenching effects may produce non-linear calibration …

Lysine 53 Acetylation Of Cytochrome C In Prostate Cancer: Warburg Metabolism And Evasion Of Apoptosis, Viktoriia Bazylianska, Hasini A. Kalpage, Junmei Wan, Asmita Vaishnav, Gargi Mahapatra, Alice A. Turner, Dipanwita Dutta Chowdhury, Katherine Kim, Paul T. Morse, Icksoo Lee, Joseph S. Brunzelle, Lisa Polin, Prabal Subedi, Elisabeth I. Heath, Izabela Podgorski, Katrin Marcus, Brian Fp Edwards, Maik HüTtemann

Biochemistry and Molecular Biology Faculty Publications

Prostate cancer is the second leading cause of cancer-related death in men. Two classic cancer hallmarks are a metabolic switch from oxidative phosphorylation (OxPhos) to glycolysis, known as the Warburg effect, and resistance to cell death. Cytochrome c (Cytc) is at the intersection of both pathways, as it is essential for electron transport in mitochondrial respiration and a trigger of intrinsic apoptosis when released from the mitochondria. However, its functional role in cancer has never been studied. Our data show that Cytc is acetylated on lysine 53 in both androgen hormone-resistant and -sensitive human prostate cancer xenografts. To characterize the …

Influence Of Monovalent And Divalent Ions In The Conformational Change Of Caspase-Cleaved Par-4 (Cl-Par-4) Tumor Suppressor Protein, Krishna K. Raut, Komala Ponniah, Steven M. Pascal

College of Sciences Posters

Prostate apoptosis response-4 (Par-4) is a pro-apoptotic tumor suppressor protein. We have shown that this 38 kDa full-length Par-4 (Fl-Par-4) protein is predominantly intrinsically disordered in vitro. In vivo, Par-4 is cleaved by caspase-3 at Asp-131 to generate a 24 kDa functionally active cleaved Par-4 (cl-Par-4) fragment. The cl-Par-4 protein inhibits the NF-κB-mediated cell survival pathway and causes selective apoptosis in various tumor cells. Our laboratory is interested in how the disorder-order balance within Fl-Par-4 and cl-Par-4 may be related to the balance between cell survival and cell death. Currently, we are using biophysical techniques such as circular …

Visible Light Generation And Mechanistic Investigation Of High-Valent Metal-Oxo Species Supported By Different Ligands, Seth Ellis Klaine

Masters Theses & Specialist Projects

Numerous transition metal catalysts have been designed as biomimetic model compounds for the active site of metalloenzymes found throughout Nature, most notably cytochrome P450 monooxygenases that carry out the oxidative transformations of organic substrates with near-perfect chemo-, regio-, and stereo-selectivity. The primary active oxidants in catalytic and enzymatic cycles are fleeting high-valent metal-oxo intermediates where the oxo ligand can transfer to an organic substrate in a process known as oxygen atom transfer (OAT).

In the present work, porphyrin-manganese(III), salen-chromium(III), and salenmanganese( III) derivatives were successfully synthesized and spectroscopically characterized using 1H NMR and UV-Vis spectroscopies. A facile photochemical approach was …

Analysis Of Student Learning Gains In A Biochemistry Cure Course During The Mandatory Covid-19 Shift To Online Learning, Ambika Kapil, Santanu De, Arthur Sikora

Biology Faculty Proceedings, Presentations, Speeches, Lectures

Due to the ongoing COVID-19 pandemic, institutions across the world have had to make modifications to existing curricula, especially in the experimental science lab. There is a need to better understand student learning in this environment. Using the Participant Perception Indicator (PPI) survey, we measure the students’ knowledge, experience, and confidence (KEC) growth over the course of a fully online biochemistry course. Using a combination of video explanations, experimental procedure documents and sample data students completed the Biochemistry Authentic Scientific Inquiry Lab (BASIL) Course-based Undergraduate Experience (CURE) in summer 2020. The results and analysis of the survey data gave light …

Computational And Experimental Investigation Into The Determinants Of Protein Structure, Folding, And Stability In The Β-Grasp Superfamily, John T. Bedford Ii

Chemistry & Biochemistry Theses & Dissertations

Elucidating the mechanisms of protein folding and unfolding is one of the greatest scientific challenges in basic science. The overarching goal is to predict three-dimensional structures from their amino acid sequences. Understanding the determinants of protein folding and stability can be facilitated through the study of evolutionarily related but diverse proteins. Insights can also be gained through the study of proteins from extremophiles that may more closely resemble the primordial proteins. In this doctoral research, three aims were accomplished to characterize the structure, folding and unfolding behavior within the β-grasp superfamily. We propose that the determinants of structure, stability, and …

Toward The Discovery Of Biological Functions Associated With The Mechanosensor Mtl1p Of Saccharomyces Cerevisiae Via Integrative Multi-Omics Analysis, Nelson Martínez-Matías, Nataliya Chorna, Sahily González-Crespo, Lilliam Villanueva, Ingrid Montes-Rodríguez, Loyda M. Melendez-Aponte, Abiel Roche-Lima, Kelvin Carrasquillo-Carrión, Ednalise Santiago-Cartagena, Brian C. Rymond, Mohan Babu, Igor Stagljar, José R. Rodríguez-Medina

Biology Faculty Publications

Functional analysis of the Mtl1 protein in Saccharomyces cerevisiae has revealed that this transmembrane sensor endows yeast cells with resistance to oxidative stress through a signaling mechanism called the cell wall integrity pathway (CWI). We observed upregulation of multiple heat shock proteins (HSPs), proteins associated with the formation of stress granules, and the phosphatase subunit of trehalose 6-phosphate synthase which suggests that mtl1Δ strains undergo intrinsic activation of a non-lethal heat stress response. Furthermore, quantitative global proteomic analysis conducted on TMT-labeled proteins combined with metabolome analysis revealed that mtl1Δ strains exhibit decreased levels of metabolites of carboxylic acid metabolism, decreased …

Construction Of E. Coli Strain With N-Terminal Domain Deletion In The Biotin Carboxyl Carrier Protein, Dara Khosravi

Honors Capstones

No abstract provided.