Biochemistry Commons^™

The Effects Of Neuronal Growth Factors On Lps-Activated Schwann Cells, Caitlyn E. Henry, Peyton Kimmel, Angela Asirvatham Ph.D.

Student Research Poster Presentations 2022

Schwann cells (SCs) are the principal support cells of neurons in the peripheral nervous system, that both myelinate axons for the rapid conduction of electrical impulses as well as assist in axonal repair during nerve injury. During nerve injury, SCs secrete tumor necrosis factor alpha (TNF-α)^1,5,6 and other proinflammatory mediators^1,6, attracting macrophages to the site of injury to induce inflammation and clear myelin debris.^1,6 Once the debris is cleared, the neuron stimulates SC proliferation by secreting neuronal mitogens, such as heregulin^3,4, and an unknown factor that activates the cAMP pathway³, an important …

Simulation Of An Inflammatory Model Using Schwann Cells, Caitlyn Henry, Peyton Kimmel, Angela Asirvatham

Student Research Poster Presentations 2022

During peripheral nerve injury, the myelin surrounding the neuronal axons is damaged, initiating an inflammatory response to remove myelin debris. Once myelin debris is cleared, Schwann cells acquire a proliferating phenotype which allows them to grow and divide so that remyelination can occur. The neuron stimulates Schwann cell division by secreting growth factors, like heregulin, and an unknown growth factor that activates the cAMP pathway. Although the role of cAMP in axonal regeneration is well-known, not much has been explored about its function in Schwann cells during nerve injury and inflammation. To simulate an inflammatory environment, the S16 Schwann cell …

Using Circular Dichroism Spectroscopy To Study Secondary Structure Of Horseradish Peroxidase: A Laboratory Experiment In Biochemistry, Shemaiah Kamran, Sarah E. Bultje, Kumar Sinniah

Summer Research

• We present a laboratory experiment in biochemistry to study the Far-UV Visible secondary structure of a protein (horseradish peroxidase, HRP) that can be adapted for a single-session laboratory experiment or a multiple-session laboratory project.
• A Circular Dichroism (CD) spectrometer provides structural information of proteins as a function of environmental changes such as pH and temperature. We present secondary structural information for the protein, horseradish peroxidase, at pH 3 and pH 7 in the temperature range 30-80° C in phosphate buffered solution (PBS).
• HRP has many applications in biochemistry and biotechnology. It is frequently used as a reporter enzyme in diagnostics …

Affinity Of Cadmium Versus Zinc For The Carbonic Anhydrase Active Site, Rachel Phan

Honors Scholars Collaborative Projects

Cadmium is a heavy metal found in tobacco plants, nickel-cadmium batteries, and as a byproduct of metal mining. The similarities between the properties of zinc ions and cadmium ions in proteins due to their 2+ charges may cause displacement of zinc by cadmium if encountered and has been shown to do so in several transcription factors. Cadmium’s ability to replace zinc- a metal found in around 3,000 human proteins- is a cause for concern, especially due to its toxicity, long half-life in the body, and larger size.

Carbonic anhydrase is an important metalloprotein protein that relies on a central zinc(II) …

The Role Of Dichloro(Ethylenediamine) Platinum Ii In Altering Cellular Iron And Copper Transport, Megan Hall

Mahurin Honors College Capstone Experience/Thesis Projects

Approximately 50% of all chemotherapy cancer patients receive a platinum chemotherapeutic during their treatment. Although these drugs are widely utilized, little is known about the cellular transport of platinum drugs. It is hypothesized that platinum utilizes transporters for other metals to enter the cell. Structural differences between compounds are predicted to affect the compounds’ ability to exploit different transporters. Also, different cell types express different transporters. Therefore, compound structure is expected to influence transport of platinum compounds differently in different cell types. This study investigated the role of the platinum drug cisplatin and platinum compound Pt(en)Cl₂ in altering iron …

Investigating Spatiotemporal Kinetics, Dynamics, And Mechanism Of Exosome Release, Anarkali Mahmood

Electronic Theses and Dissertations

Exosomes are small lipid-based vesicles that can carry biomolecules from one cell to another. While exosomes are crucial to maintain homeostasis in healthy cells, they are exploited by unhealthy cells to aid disease progression. Exosomes likely facilitate disease progression via the transfer of disease-causing biomolecules from unhealthy to healthy cells. Exosomes are generated in Multivesicular endosomes (MVEs) and are then secreted into the extracellular space to travel to other cells. Despite being a crucial step, very little is known about exosomes release mechanism and dynamics. To further our understanding of exosomes, specifically their secretion, my work has focused on investigating …

Radiation Exposure Determination In A Secure, Cloudbased Online Environment, Ben C. Shirley, Eliseos J. Mucaki, Joan H.M. Knoll, Peter Rogan

Biochemistry Publications

Rapid sample processing and interpretation of estimated exposures will be critical for triaging exposed individuals after a major radiation incident. The dicentric chromosome (DC) assay assesses absorbed radiation using metaphase cells from blood. The Automated Dicentric Chromosome Identifier and Dose Estimator System (ADCI) identifies DCs and determines radiation doses. This study aimed to broaden accessibility and speed of this system, while protecting data and software integrity. ADCI Online is a secure web-streaming platform accessible worldwide from local servers. Cloud-based systems containing data and software are separated until they are linked for radiation exposure estimation. Dose estimates are identical to ADCI …

Kinase-Catalyzed Labeling To Identify Kinase-Substrate Pairs Using Γ-Phosphate Modified Atp Analogs, Rachel Beltman

Wayne State University Dissertations

Post-translational modifications (PTMs) are responsible for a variety of cellular processes. One such PTM is protein phosphorylation, which is catalyzed by kinases. Kinase enzymes play important roles in cellular signaling pathways, but dysregulation of kinase-mediated events results in the formation of diseases, which make kinases favorable drug targets. To uncover the role kinases play in the development of diseases, kinase-mediated cellular events need to be better understood. The current gap in the field is the lack of tools available to identify the kinase that is responsible for specific phosphorylation events within the cell. To improve the gap in the field, …

Metabolic Synergy Between Human Symbionts Bacteroides And Methanobrevibacter, Jennie L. Catlett, Sean Carr, Mikaela Cashman, Megan D. Smith, Mary Walter, Zahmeeth Sakkaff, Christine A. Kelley, Massimiliano Pierobon, Myra B. Cohen, Nicole R. Buan

Department of Biochemistry: Faculty Publications

ABSTRACT Trophic interactions between microbes are postulated to determine whether a host microbiome is healthy or causes predisposition to disease. Two abundant taxa, the Gram-negative heterotrophic bacterium Bacteroides thetaiotaomicron and the methanogenic archaeon Methanobrevibacter smithii, are proposed to have a synergistic metabolic relationship. Both organisms play vital roles in human gut health; B. thetaiotaomicron assists the host by fermenting dietary polysaccharides, whereas M. smithii consumes end-stage fermentation products and is hypothesized to relieve feedback inhibition of upstream microbes such as B. thetaiotaomicron. To study their metabolic interactions, we defined and optimized a coculture system and used software testing …

Arginine Catabolism And Polyamine Biosynthesis Pathway Disparities Within Francisella Tularensis Subpopulations, Yinshi Yue, Bhanwar Lal Puniya, Tomáš Helikar, Benjamin Girardo, Steven H. Hinrichs, Marilyn A. Larson

Department of Biochemistry: Faculty Publications

Francisella tularensis is a highly infectious zoonotic pathogen with as few as 10 organisms causing tularemia, a disease that is fatal if untreated. Although F. tularensis subspecies tularensis (type A) and subspecies holarctica (type B) share over 99.5% average nucleotide identity, notable differences exist in genomic organization and pathogenicity. The type A clade has been further divided into subtypes A.I and A.II, with A.I strains being recognized as some of the most virulent bacterial pathogens known. In this study, we report on major disparities that exist between the F. tularensis subpopulations in arginine catabolism and subsequent polyamine biosynthesis. The genes …

Principles Of General Chemistry, Jose Cobo

Open Educational Resources

No abstract provided.

Investigation Of Catalysis Of Nitration By Cytochrome P450s, Lannika Johnson

Honors Undergraduate Theses

TxtE is a protein related to cytochrome P450 enzymes, which catalyze a number of reactions that typically involve oxygen and not nitrogen. It has been discovered that TxtE can nitrate tryptophan through an unusual reaction in which it uses nitric oxide (NO) as a nitrogen donor to install the nitro group despite NO typically being considered toxic to bacteria. This project will determine if all cytochromes P450 can catalyze nitration as long as they are given NO. This will have an impact on understanding drug delivery and metabolism for which nitration is important.

Examining The Effects Of Exercise-Induced, Physical Stress Overtraining On Stress Biomarkers In Adolescent, C57bl/6 Mice, Curtis Scotty Davis

Theses, Dissertations and Capstones

Exercise has long been established as a therapeutic modality to improve health and related physical fitness, sports performance, and injury/risk reduction in both adults and adolescents (Haff & Triplett, 2015; Kaminsky, 2010). Nonetheless, there is a lack of research investigating the negative effects of exercise overstress (i.e., overtraining syndrome) in these populations, particularly adolescents (Brenner & American Academy of Pediatrics Council on Sports Medicine and Fitness, 2007; Matos & Winsley, 2007). The objective of this study was to examine hormone biochemical markers, anthropometric markers, and behavioral traits, which are associated with overtraining syndrome, in adolescent C57BL/6 mice. A total of …

Complex Dna Mixture Deconvolution By Single Cell Genomics: Direct Physical Cell Subsampling And High Sensitivity Dna Typing, Kaitlin Huffman

Electronic Theses and Dissertations, 2020-2023

Biological evidence is often found at crime scenes and can comprise of DNA from the victim(s) and perpetrator(s) to a crime as well as from individuals with no direct relationship to the incident. This can complicate analysis as DNA mixtures are one of the more difficult sources of biological evidence to interpret. Probabilistic genotyping (PG) has greatly aided in mixture analysis. However, even with PG, standard bulk mixture approaches do not always result in probative results as allele overlap, artifacts, or low-level minor contributors inevitably cause genotype information loss. Therefore, deconvolution of forensic DNA mixtures into their individual component DNA …

G-Quadruplex Reporters: Structural Studies And Application For Visual And Fluorescent Detection Of Point Mutations In Nucleic Acid Sequences, Ryan Connelly

Electronic Theses and Dissertations, 2020-2023

DNA-based diagnostics traditionally utilize hybridization probes: strands of DNA complimentary to a target sequence that, upon binding, generate a signal to indicate the presence of the target. The classic hybridization probes are the molecular beacon (MB) and TaqMan probes, both single DNA strands with a fluorophore and quencher at opposing ends. Despite their widespread use in applications such as qPCR due to their ability to multiplex with a variety of bound fluorophores, these probes have several shortcomings: temperature limitations for selective target recognition and differentiating single-nucleotide polymorphisms, intrinsic design limitations to interrogate some target sequences, and a high relative cost …

Harnessing The Power Of The Nitro Group In Natural Product Synthesis, Katelyn Bobek

Electronic Theses and Dissertations, 2020-2023

The high reactivity and electron-withdrawing ability of the nitro group makes it a powerful tool in synthesizing a diverse range of bioactive natural products. The potential of the nitro group was employed via two main methodologies: iron (III)-mediated oxidative dearomatization and oxime carbonate butyrolactone formation to reach three important heterocyclic scaffolds; spiro, tropone and γ-butyrolactone. Nitro precursors were enantioselective synthesized by key nitro-aldol (Henry) and nitro-Michael reactions. A library of polysubstituted butyrolactone oxime carbonates were formed under mild DMAP and Boc2O conditions from readily available δ-nitroalcohols in good to high yields with broad functional group tolerability. Six lignan γ-butyrolactone natural …

Extending Plasma Proteome Coverage To Enable Biomarker Discovery, Jordan Bruce Burton

Wayne State University Dissertations

Two features make plasma a valued source of health information: First, plasma contains secreted proteins from all tissues in the body and can, therefore, provide information about the health status of all tissues. Second, plasma is available in large quantity using a minimally invasive procedure that is well tolerated. The status of plasma as a valued source of health information is matched by the difficulty in extracting proteome information from the samples. Using state of the art instrumentation a standard analysis of plasma yields quantitative information on approximately 300 proteins. Extreme fractionation strategies coupled with increased mass spectrometry time extends …

Synthesis Of A Key Building Block For A Metal-Like Carbon-Based Compound, Chimi Dolker Sherpa

Senior Honors Theses and Projects

No abstract provided.

X-Ray Crystallography Of Protein Uhrf2 With H3k9me3, Isaak Miller

Senior Honors Theses and Projects

Epigenetics is the study of regulating gene expression without changing the DNA sequence. Ubiquitin-like containing Plant Homeodomain and Ring-Finger 2 (UHRF2) is an epigenetic protein that is both overexpressed and underexpressed in various types of cancers. Recent studies have shown the Tandem Tudor Domain (TTD) and Planthomeo Domain (PHD) of UHRF2 interact with histone 3 trimethylated at lysine 9 (H3K9me3). However, the structural basis for UHRF2 TTD-PHD binding H3K9me3 has not been determined. This project consists of purification of the UHRF2 TTD-PHD protein construct expressed in E. coli, crystallization of the protein, X-ray diffraction of the crystal, and molecular replacement …

Functionalization Of Chitosan Based Microparticles For In Vitro 3d Culture Of Human Liver Cells, William Imes

Williams Honors College, Honors Research Projects

Previous work involving 3D culture of human liver cells with fluorinated chitosan based microparticles has shown that their incorporation provides needed structural cues that the culture of cells alone does not provide, such as increased gas transport. However, using bare microparticles to grow large 3D cellular structures is not practical as they tend to collapse before any meaningful research can be done on them. It is proposed that this is because of the lack of extracellular matrix (ECM)components within pure cell culture in vitro, which in their absence cannot adequately facilitate biochemical communication and adhesion between cells. It is …