Biochemistry, Biophysics, and Structural Biology Commons^™

Study Of Protein Extraction Techniques For M. Foliorum Phages, Lola Sibaud, Anna Elgersma

Student Academic Conference

Characterizing phages is becoming increasingly important in biochemistry and biotechnology due to their potential to treat bacterial infections without need for antibiotics. Last year, students attempted to isolate proteins from Microbacterium foliorum infecting phages Babydotz and Rosadiaz. Proteins were successfully extracted; however, after further analysis it was shown that the obtained proteins belonged to the host bacteria and not the phages. The purpose of this study was to find a method to separate phage from host bacterial debris to successfully isolate phage proteins and analyze them later. Isolating phage proteins could help us characterize more phages and know more about …

Novel Plant Peroxidase Investigation For Use In Various Biochemical And Practical Applications, Sally Hess

Liberty University Research Week

Undergraduate

Basic

Activation Of Nrf2 Pathway By Natural Products, Genesis Schat, Madelyn Sedlachek

Liberty University Research Week

Undergraduate

Basic

Characterization Of A Sas3 Homolog In The Human Pathogenic Yeast Cryptococcus Neoformans, Lillianne Bartone

Liberty University Research Week

Undergraduate

Basic

Characterization Of Select Lysine Mutations Of The Cystine/Glutamate Transporter, System XC-, Anna Koppin, Claire Buck, Amanda Gibson

22nd Annual Celebration of Undergraduate Research and Creative Activity (2023)

System xc- is a membrane transport system that plays a critical role in mitigating oxidative stress. As such, its regulation is critical for proper brain functioning. Recent work in our lab has shown that System xc- activity increases immediately during an oxidative insult by undergoing a change in localization to the plasma membrane, but we have yet to identify the specific mechanism for the redistribution of the transporter. Previous studies have demonstrated that post-translational modifications of proteins can lead to differential protein distribution within cells. Therefore, in this study, we sought to determine if post-translational modification (PTM) of the transporter …

Does Phosphorylation On Serine 26 Of System Xc- Lead To Changes In Cell Surface Expression?, Katherine Lane

22nd Annual Celebration of Undergraduate Research and Creative Activity (2023)

System x_c^- is involved in transporting cystine into cells and glutamate out of cells, and ultimately in production of the antioxidant glutathione. Antioxidants are important in protecting cells from oxidative stress which can occur when waste products like H₂O₂ build up in the cell. Previous studies have shown that a critical regulator mTORC regulates system xc- by phosphorylating serine 26 (S26) on the N-terminus of the cystine/glutamate antiporter xCT, leading to a reduction in transport activity. The specific objective of this study is to determine the mechanism by which phosphorylation of S26 affects activity. To …

Creation Of An N-Terminal Xct Mutant Lacking Lysines For Use In Protein Turnover Studies, Alexandria Switzer

22nd Annual Celebration of Undergraduate Research and Creative Activity (2023)

xCT plays a role in protecting cells from oxidative stress as well as intracellular glutathione synthesis. If this process is impeded, reactive oxygen species accumulate in the cell, leading to cellular damage, and if left unchecked, it can lead to neuronal loss. Ubiquitin is a small protein that some evidence suggests can negatively affect the stress response pathway modulated by xCT, as well as induce cell death. It is hypothesized that ubiquitin binds to xCT on the N-terminal lysine residues, and there are six conserved lysines in the N-terminal domain at positions K4R, K12R, K30R, K37R, K41R, and K43R. Previous …

Early Life Δ9-Tetrahydrocannabinol Exposure Of F0 Zebrafish Causes Hyperactivity In F1 Offspring, Jenna Cripe

Honors Theses

As cannabis increases in its accessibility, potency, and acceptance across the United States, investigation into the multigenerational effects of Δ9-tetrahydrocannabinol (THC) exposure during key stages of development is critical. The aim of this study was to evaluate whether there would be behavioral impacts in the F1 offspring following a dose-response of THC exposure (0.08, 0.4, or 1 µM) during development in the F0 generation. Zebrafish (Danio rerio) were utilized in this study. Behaviors (locomotive activity and anxiety-like behavior) in the F1 generation were evaluated at 120 hours post-fertilization (hpf) with the larval photometer response (LPR) assay, and at 3, 11, …

A Fisher Indole Synthesis Approach To Phidianidine Analogues, Anna Tingler, Trinity Ghering, Samuel Ross

Undergraduate Research Competition

Naturally occurring products are molecules that are found in the natural world without the aid of humans. Aspirin and opium are famous examples of this classification of molecule. Another example of natural products that are less known are the phidianidines. Phidianidines A & B were isolated from a shell-less mollusk, Phidiana militaris. These molecules are comprised of a 1,2,4-oxadiazole ring, an indole and an aminoalkylguanadio group. These compounds have exhibited striking pharmacological behavior such as that of neutralizing reactive oxygen species and acting as an agonist for the μ-opioid receptor. In recent years, the synthesis of phidianidine analogues has replaced …

A New Approach To The Core Of Flinderole C, Ana Pettijohn, Claire Romain

Undergraduate Research Competition

The flinderoles A-C are a class of anti-malaria bisindole alkaloids from plants of the Flindersia genus. Flinderole C was specifically from the Papua New Guinean plant called F. amboinensis and is the most potent antimalarial of the group. Malaria is a parasitic infectious disease usually found in the hot, tropic regions of the world, such as parts of Africa. It is estimated that nearly half the world's population lives in an area where it is endemic. Furthermore, malaria was estimated to have caused over 600,000 deaths in 2020. While measures to limit the spread of the disease lessen its impact, …

Discovery And Characterization Of A Prophage Encoded Anti-Crispr Against The Type Iv-A Crispr System, Andrew Williams

Student Research Symposium

CRISPR Systems are adaptive immune systems in bacteria used to defend against mobile genetic elements such as bacteriophages, or bacterial viruses, and plasmids. They use a protein complexed with a guide RNA to target the protein to a foreign invader and neutralize it by cleaving its genetic information. In order to survive and keep infecting bacteria, bacteriophages likewise have developed machinery used to inhibit CRISPR immunity in the form of anti-CRISPR (Acr) proteins. The type IV-A system is a recently discovered and characterized system by the Jackson lab. It has been shown to defend against plasmids, but further data on …

Cycloaddition Of Aryldiazoacetates With Terminal Alkynes And Nitriles Via A Bioinspired Cobalt Complex, Liming Tan

Student Research Symposium

In this study, we present a novel method for the efficient cycloaddition of aryl diazoacetates with both terminal alkynes and nitriles, employing a bioinspired cobalt complex as a catalyst. Our findings indicate that terminal alkynes react with aryl diazoacetates to yield a cyclopropenation product through a 2+1 cycloaddition reaction. Meanwhile, nitriles react with aryl diazoacetates to produce an oxazole product via a 2+3 cycloaddition reaction. These results have significant implications for the development of new synthetic routes towards cyclopropenation and oxazole compounds.

Developing A Safe And Effective Papillomavirus Screen To Be Used On College Students, Nicole Frantz

Undergraduate Research Competition

Human papillomavirus (HPV) is the most common sexually transmitted infection that accounts for approximately 5% of all cancers worldwide and affects more than 80 million people in the US alone, according to the CDC and National Cancer Institute. Human papillomaviruses are small, nonenveloped, icosahedral DNA viruses that infect squamous epithelial cells. The viral particles consist of a single double stranded DNA molecule bound to histones and contained within a protein capsid composed of structural proteins late (L)1 and L2. To date, over 100 different genotypes of HPV have been identified, and approximately 15 types are considered oncogenic in cervical, vulvar, …

Environmental Bacteriophage Presence In The Drainage Ponds At Coastal Carolina University, Alexei Chesnutwood

Undergraduate Research Competition

The purpose of this research is to detect naturally occurring, lytic bacteriophages and identify the environmental factors that influence their presence on Coastal Carolina University’s campus. Bacteriophages are non-living viruses that only infect bacteria, and these viruses are found in abundance in every environment. Likewise, coliphages are viruses that exclusively infect coliform bacteria. Eleven treated and untreated freshwater sample sites were chosen for weekly sampling on CCU’s campus. During water sample collection, ambient environmental conditions were measured. In addition, precipitation was tracked over the collection period as runoff relates to the proliferation of bacteriophages. If plaque assays indicated the presence …

Toward Quantification Of Hypoxia Using Fluorinated Euii/Iii-Containing Ratiometric Probes, S A Amali S Subasinghe, Caitlyn J Ortiz, Jonathan Romero, Cassandra L Ward, Alexander G Sertage, Lyazat Kurenbekova, Jason T Yustein, Robia G Pautler, Matthew J Allen

Faculty, Staff and Students Publications

Hypoxia is a prognostic biomarker of rapidly growing cancers, where the extent of hypoxia is an indication of tumor progression and prognosis; therefore, hypoxia is also used for staging while performing chemo- and radiotherapeutics for cancer. Contrast-enhanced MRI using EuII-based contrast agents is a noninvasive method that can be used to map hypoxic tumors, but quantification of hypoxia using these agents is challenging due to the dependence of signal on the concentration of both oxygen and EuII. Here, we report a ratiometric method to eliminate concentration dependence of contrast enhancement of hypoxia using fluorinated EuII/III-containing probes. We studied three different …

A Ferritin Nanoparticle-Based Zika Virus Vaccine Candidate Induces Robust Humoral And Cellular Immune Responses And Protects Mice From Lethal Virus Challenge, Aryamav Pattnaik, Bikash R. Sahoo, Lucas R. Struble, Gloria E. O. Borgstahl, You Zhou, Rodrigo Franco, Raul G. Barletta, Fernando A. Osorio, Thomas M. Petro, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

The severe consequences of the Zika virus (ZIKV) infections resulting in congenital Zika syndrome in infants and the autoimmune Guillain–Barre syndrome in adults warrant the development of safe and efficacious vaccines and therapeutics. Currently, there are no approved treatment options for ZIKV infection. Herein, we describe the development of a bacterial ferritin-based nanoparticle vaccine candidate for ZIKV. The viral envelope (E) protein domain III (DIII) was fused in-frame at the amino-terminus of ferritin. The resulting nanoparticle displaying the DIII was examined for its ability to induce immune responses and protect vaccinated animals upon lethal virus challenge. Our results show that …

Moderate-Intensity Exercise Improves Mesenteric Arterial Function In Male Uc Davis Type-2 Diabetes Mellitus (Ucd-T2dm) Rats: A Shift In The Relative Importance Of Endothelium-Derived Relaxing Factors (Edrf), Md Rahatullah Razan, Said Amissi, Rifat Ara Islam, James L. Graham, Kimber L. Stanhope, Peter J. Havel, Roshanak Rahimian

School of Pharmacy Faculty Articles

The beneficial cardiovascular effects of exercise are well documented, however the mechanisms by which exercise improves vascular function in diabetes are not fully understood. This study investigates whether there are (1) improvements in blood pressure and endothelium-dependent vasorelaxation (EDV) and (2) alterations in the relative contribution of endothelium-derived relaxing factors (EDRF) in modulating mesenteric arterial reactivity in male UC Davis type-2 diabetes mellitus (UCD-T2DM) rats, following an 8-week moderate-intensity exercise (MIE) intervention. EDV to acetylcholine (ACh) was measured before and after exposure to pharmacological inhibitors. Contractile responses to phenylephrine and myogenic tone were determined. The arterial expressions of endothelial nitric …

Locomotor Biomechanics In Stream Gobies, Rhinogobius Sp.: Functional Performance Of Waterfall-Climbing Behavior, Ernest W. Murphy

Student Scholar Showcase

Gobiidae is the taxonomic classification of a large fish family comprising over 2,000 species. A few goby species have the unique ability of scaling waterfalls using pelvic fins that are fused into a suction disc (pelvic sucker) located on the ventral side of the body, aiding them in upstream migration to freshwater feeding and reproduction locations. This study aimed to investigate the locomotor biomechanics of Rhinogobius goby fish due to the lack of literature in the area. In particular, climbing performance variables (pressure differential, force, endurance, and kinematics of attachment), pelvic sucker fatigability, and ontogenetic patterns of physical development and …

Analysis Of Attitudinal Student Learning Benefits From A Course-Based Undergraduate Research Experience (Cure) Adapted For Online Format, Ambika Kapil, Luis C. Gonzalez Isoba, Niraj Pathak, Arthur Sikora, Santanu De

FDLA Journal

Course-based Undergraduate Research Experiences (CUREs) are an increasingly utilized model for exposing students to research. The lack of robust assessments is a major hurdle to wider adoption of CUREs. The Coronavirus Infectious Disease 2019 (COVID-19) pandemic necessitated a drastic shift of in-person courses to the online format. Using the Participant Perception Indicator (PPI) survey, we measured students’ self-reported changes in learning from such a biochemistry course at a large university in south Florida based on the Biochemistry Authentic Scientific Inquiry Lab (BASIL) model. By doing this, we were able to better understand the student-benefits of CUREs and how these benefits …

The Global Transcription Factor Fur Is Activated By Binding A [2fe-2s] Cluster To Maintain Iron Homeostasis In Escherichia Coli, Chelsey Rene Fontenot

LSU Doctoral Dissertations

Iron is an essential element for biological functions. Organisms have developed specific mechanisms to acquire, store, and export iron to manage intracellular iron homeostasis. These mechanisms are crucial not only for providing the cells with adequate iron for biological processes but also for ensuring that cells are not overloaded with excess iron, which is highly toxic. The Ferric uptake regulator (Fur) is a global transcription factor for maintaining intracellular iron homeostasis in bacteria. Previously, it was postulated that when the intracellular iron content is elevated, Fur binds ferrous iron to down-regulate the iron acquisition genes and up- regulate the iron …

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

Masters Theses

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

Characterizing Marr Homologs That Mediate Bacterial Fitness And Survival, George Chukwuemeka Nwokocha

LSU Doctoral Dissertations

Plant disease outbreaks are on the rise, threatening food security in many parts of the globe. To address these enormous challenges, new intervention strategies like bio-controls are being used as an important sustainable alternative to mitigate the negative effects of pathogens and increase the quality and productivity of agricultural crops. MarR family transcriptional regulators have been studied for their involvement in regulating bacterial virulence genes that help colonize plant hosts and therefore could serve as targets for biocontrol. The transcriptional regulator PecS, a member of the MarR family, is encoded by select bacterial pathogens and we show here that PecS …

The Effects Of Specialized Pro-Resolving Mediator Lipoxin A4 On Pseudomonas Aeruginosa Biofilms And Interactions With Monocytes, Julianne M. Thornton

Graduate School of Biomedical Sciences Theses and Dissertations

Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen known as a major cause of hospital-acquired secondary infections, commonly causing chronic respiratory infections in immunocompromised individuals, especially those with cystic fibrosis, and often found in wound infections. P. aeruginosa uses the quorum sensing pathway to readily form protective biofilms, which reduce the efficacy of antibiotics and access by host immune cells to eradicate the pathogen. Specialized pro-resolving mediators (SPMs) are lipids endogenously produced by the host immune response to infection to aid in infection resolution. One SPM, Lipoxin A4 (LxA4), has been shown to be a robust quorum sensing inhibitor.

The …

Modeling The Tripartite Role Of Cyclin C In Cellular Stress Response Coordination, Steven J. Doyle

Graduate School of Biomedical Sciences Theses and Dissertations

For normal cellular function, exogenous signals must be interpreted and careful coordination must take place to ensure desired fates are achieved. Mitochondria are key regulatory nodes of cellular fate, undergoing fission/fusion cycles depending on the needs of the cell, and help mediate cell death fates. The CKM or Cdk8 kinase module, is composed of cyclin C (CC), Cdk8, Med12/12L, and Med13/13L. The CKM controls RNA polymerase II, acting as a regulator of stress-response and growth-control genes. Following stress, CC translocates to the mitochondria and interacts with both fission and iRCD apoptotic mediators. We hypothesize that CC represents a key mediator, …

Investigating The Antibacterial And Immunomodulatory Properties Of Lactobacillus Acidophilus Postbiotics, Rachael M. Wilson

Graduate School of Biomedical Sciences Theses and Dissertations

Probiotics are nonpathogenic microorganisms that have been extensively studied for their ability to prevent various infectious, gastrointestinal, and autoimmune diseases. The mechanisms underlying these probiotic effects have not been elucidated. However, we and other researchers have evidence suggesting that probiotic bacteria secrete metabolites that are antimicrobial and anti-inflammatory. As such, we developed a methodology to collect the secreted metabolites from a probiotic bacterium, Lactobacillus acidophilus, and tested this cell free filtrate (CFF) both in vitro and in vivo. Using this CFF, we have demonstrated that L. acidophilus secretes a molecule(s) that has specific bactericidal activity against the opportunistic pathogen, Pseudomonas …

Analysis Of Ssa4 Reporter Expression By Q-Pcr, Susveen Sharanshi, Rebecca Adams

[Archive] Belmont University Research Symposium (BURS)

The synthesis of genome-encoded proteins via mRNA translation is integral to cell survival. In eukaryotes, such as S. cerevisiae, the mRNA that is produced in the nucleus must be exported to the cytoplasm for translation to occur, and this process is highly regulated. Specifically, the export of mRNA occurs via travel through nuclear pore complexes (NPCs), which are selective doorways embedded in the nuclear envelope. During cellular stress, such as heat shock, the cell needs to regulate gene expression to permit survival, and mRNA export is one step at which this occurs. At these high temperatures, a cell’s proteins …

Med13 Degradation Defines A New Receptor-Mediated Autophagy Pathway Activated By Nutrient Deprivation, Sara E. Hanley

Graduate School of Biomedical Sciences Theses and Dissertations

Cells are exposed to an enormous amount of diverse extracellular cues but have a limited arsenal of weapons for protecting and maintaining homeostasis. To overcome these restrictions, nature has engineered proteins that have multiple functions. The pleiotropy of using one protein to carry out a variety of functions allows cells to rapidly execute tailored responses to a diverse set of signals. The Cdk8 kinase module (CKM) is a conserved detachable unit of the Mediator complex predominantly known for its role in transcriptional regulation. The CKM is composed of four proteins, the scaffolding proteins Med13 and Med12, as well as the …

Serving Justice Case By Case, Michael Becker

WRIT: Journal of First-Year Writing

Forensic Science is an integral part of serving justice in today's society, and majority of the world has little understanding of what forensics is. Read this article on forensic science and how it relates to writing in today's world.

Physics 422 Spring 2023 Syllabus, Ronald Koder

Open Educational Resources

Spring 2023 course syllabus for Physics 422, Biophysics, at the City College of New York

Investigation Of Cofactor Activities Of Endothelial Microparticle- Thrombomodulin With Liposomal Surrogate, Valentinas Gruzdys, Lin Wang, Dan Wang, Rachel Huang, Xue-Long Sun

Chemistry Faculty Publications

Thrombomodulin (TM) is a type I transmembrane glycoprotein mainly expressed on the endothelial cells, where it binds thrombin to form the thrombin-TM complex that can activate protein C and thrombin-activable fibrinolysis inhibitor (TAFI) and induce anticoagulant and anti-fibrinolytic reactions, respec-tively. Cell activation and injury often sheds microparticles that contain membrane TM, which circulate in biofluids like blood. However, the biological function of circulating microparticle-TM is still unknown even though it has been recognized as a biomarker of endothelial cell injury and damage. In comparison with cell membrane, different phospholipids are exposed on the microparticle surface due to cell membrane "flip-flop"upon …