Biochemistry, Biophysics, and Structural Biology Commons^™

Effect Of An Oncogenic Her2 Mutant On Egfr Activity, Paige Kalik

Williams Honors College, Honors Research Projects

Studies have shown that HERs are involved in the progression of human cancers. Researchers have found that HER2 mutations in colorectal cancer are similar to mutations found in breast cancer. Recombinant antibodies are effective therapeutic treatments for cancers. Antibodies can also be used to determine how proteins function under normal and pathophysiological conditions while also identifying interacting partners. In this Honors Project, my aim is to investigate the effect of an oncogenic mutant of HER2. Considering the information above, the question researched in this project was: how does the S310F mutation affect the organization and function of HER2? The specific …

The Affect Of Dha Concentration On Lipid Membrane Fluidity, Elise Rezabeck

Williams Honors College, Honors Research Projects

My proposed project will be creating large unilaminar vesicles with varying concentrations of DHA in the vesicles. The diffusion in and out of these vesicles will be tracked by using singular lipid molecules. These singular lipid molecules will be tagged with fluorescence and viewed using an internal reflection fluorescence microscope. The data collected over the course of this project will then be analyzed using Matlab. The result of the project will be to find the specific lipid diffusion constant of these unilaminar vesicles. The relationship between the concentration of DHA and lipid diffusion and membrane fluidity will be found as …

Comparison Of Bone Robustness Between Fore- And Hindlimb In Neonatal And Infant Galago, Jeremy Busken

Williams Honors College, Honors Research Projects

According to Wolff’s law, bone is constantly subject to change based on loading patterns (Ruff et al., 2006). Those subjected to greater stress become more robust by increasing bone mineral density and often changing geometric properties. Previous studies of primate anatomy have found significant differences in bone robustness between the fore- and hindlimbs in adult leaping primates. Because leaping primates, such as Galago senegalensis and Galago moholi, subject their hindlimbs to much greater stress, this difference in robustness may come as a result of bone functional adaptation/Wolff’s law. This study sought to investigate this possibility by testing neonatal and infant …

Effects Of Nicotine Vs Non-Nicotine E-Cigarettes On The Aggression Behavior Of Crayfish, Alex Kimble

Williams Honors College, Honors Research Projects

This study looks at the effects of e-cigarette vapor with and without nicotine on the aggression behavior of crayfish (Procambarus clarkii). The treatment groups were control, nicotine, and non-nicotine. The mass, sex, and condition of their appendages were recorded before random assignment. For the purpose of this study only male crayfish were used as there were not enough female crayfish. The crayfish were then randomly assigned to each group and contained ten crayfish that were subjected to the e-cigarette vapor infused water environment for a twenty-four hour period. The control group did not contain e-cigarette vapor and was exposed to …

Biodegradation Of Rubber Particles In Soil: Using Acclimated Bacteria Isolated From Kansas Soil To Degrade Cryogrinds In Slurry, Shane Graham

Williams Honors College, Honors Research Projects

This study investigated the viability of bioremediating rubber cryogrind using enriched indigenous bacteria. To begin the experiment, soils from three highway roadside locations in Kansas, KS 96 and West, KS 400 and 143^rd, and 199^th, were collected and transported to the lab to be studied. An initial soil characterization was run on the soil samples using distilled (DI) water mixture and 0.01 M CaCl₂ to assess conductivity. The soils were tested to gather a baseline of the relationship between pH and conductivity and the impact of its distance from the roadside. Bacteria were isolated from …

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 …

Mimicking The Namib Desert Beetle With Hydrophilic And Hydrophobic Surfaces, Tyler Bracewell

Williams Honors College, Honors Research Projects

The Namib Desert beetle, Stenocara gracilipes, condenses fog that rolls in off the west coast of Namibia to provide itself with the water it needs for the day. By lifting its abdomen into the air to collect the water vapor, it condenses on hydrophilic bumps that are about 500μm in diameter and are spaced apart about 500-1500μm. Once the water reaches a critical size, the droplet rolls off the hydrophilic bump and onto the flat hydrophobic surface of the elytra. Since the beetle is angled with its abdomen in the air, the water rolls down to its mouth. A …

Sequential Block Copolymers Of Polyisobutylene And Polyolefins, Manwel Labib

Williams Honors College, Honors Research Projects

A two-step approach will be utilized to synthesize random polyolefin multiblock-copolymers. This strategy takes advantage of a,w-terminated telechelic polyolefins. Polypropylene (iPP)/polyethylene (PE) are picked because of their strength, and polyisobutylene (PIB) because of its hemocompatibility. This copolymer is theorized to make for robust bio-inert plastic, which can be used for surgical mesh implants.

The Influence Of Hydrogen Peroxide On The Enrichment Of Fe(Iii) Reducing Bacteria From Acid Mine Drainage, Susami Seth

Williams Honors College, Honors Research Projects

It is hypothesized that the ocean of Europa, a Jupiter moon, hosts bacteria on its oceanic floor. Understanding how Fe(III) reducing bacteria (FeRB) from AMD utilize organic materials within its surrounding environment outlines how FeRB could thrive and tolerate extreme conditions. FeRB are known to tolerate metals and highly reactive oxidants species (ROS), but in this experiment, H₂O₂ was the experimental factor to further test FeRB tolerance. H₂O₂ is a common ROS and is damaging to living material such as proteins, DNA, and RNA. A range of H₂O₂ concentrations were fed …

Synthesis Of Rna Nucleotides Under Probable Prebiotic Conditions, Ryan Stimson

Williams Honors College, Honors Research Projects

RNA being composed of multiple covalently linked nucleotides is thought to have been a precursor to life circa 4.3-3.8 billion years ago. Non-enzymatically formed adenosine monophosphate (AMP), more specifically, is a vitally important subtopic of the self-assembly of the first RNA sequence. The goal of this study was to synthesize AMP non-enzymatically under benign conditions that are likely to have existed on early Earth. In this experiment, 3’,5’-cAMP was successfully formed using wet-dry cycles at 80°C paired with the minerals zeolite beta, hydroxyapatite, and aerosil 300 in the presence of adenosine, urea, and pyrophosphate. A nuclear magnetic resonance spectrometer was …

Titanium Dioxide Nanoparticles Binding To Human Blood Plasma Proteins, Morgan Miller

Williams Honors College, Honors Research Projects

Titanium Dioxide (TiO₂, titania) nanoparticles are widely studied in cosmetology, food administration, and medical fields. This is due to the nanoparticles’ suitable properties, specific combination of strength and high corrosion resistance. With the increasing attention to these particles, there is concern with potential health complications. Some of the complications involved with titania nanoparticles include blood clots and immune reactions (Sit et al., 2019). In this study, we continue the study conducted in the Sahai research group (J. Chen. MS thesis, 2020), on the binding of titania nanoparticles with blood plasma proteins, focusing on potential effects of shape only. …

The Effects Of Increasing Positively Charged Metal Ions Within Synovial Fluid, Kandisi Anyabwile

Williams Honors College, Honors Research Projects

Osteoarthritis is a degenerative joint disease that affects 10% of men and 13% of women over age of 60. It is the degradation of the cartilage between two bones; obesity, age, overuse, or injury are major contributors to the development of this disease. The joint is incapsulated by the synovial sac filled with a viscous solution that aids in lubrication referred to as synovial fluid. If the synovial sac is ruptured due to injury, positive ions (K⁺, Na⁺, Ca²⁺, and Fe³⁺) may affect viscoelastic properties within the sac. The purpose of this …

The Effect Of Cxcl12 Ligand On Internalization And Dimerization Of Cxcr4 Receptors In Live Cells, Loga Iyer

Williams Honors College, Honors Research Projects

The primary objective of this project was to determine the effect of CXCL12 ligand binding on the CXCR4 receptor, specifically, how it would impact receptor internalization and dimerization. The CXCL12 ligand derives from the stromal cell-derived alpha family [8]. The CXCR4 receptors, known as C-X-C chemokine receptor type 4 play an essential role in controlling cell proliferation. When misregulated, these receptors can drive tumorigenesis and are thus important targets of cancer therapy. These G protein-coupled receptors stimulate a cascade of signaling pathways in specific tissues [1]. These pathways include the positive transcriptional control of CXCR4 via the Nuclear Respiratory Factor-1 …

Non-Invasive Method For Leptin Supplementation In Zebrafish (Danio Rerio), Regan Mcnamara

Williams Honors College, Honors Research Projects

I tested the hypothesis that recombinant leptin protein can be introduced to zebrafish in vivo through non-invasive soaking in a solution containing the protein. One way to study various molecules’ effects in vivo is through intraperitoneal or intracerebroventricular injections during the embryonic or larval stage, which is invasive, difficult to administer, and can have a high mortality rate. 48 hours post fertilization (hpf) zebrafish were soaked in a His-tagged recombinant leptin protein solution at 10 nM and 100 nM concentrations (produced by Genscript). After soaking, zebrafish larvae were washed extensively to remove all recombinant protein on their exterior before homogenization. …

Synthesis, Characterization, And Biological Activity Of Imidazolium Salts, David Weader

Williams Honors College, Honors Research Projects

Nonmuscle invasive bladder cancer (NMIBC) inflicts thousands of Americans annually, and is typically treated with the immunotherapy BCG. However, due to a BCG shortage, there is a new need for novel treatments of NMIBC. Addressing this issue, several imidazolium salt derivatives were synthesized and characterized with the intent of treatment within the bladder. These imidazolium salts were tested against different human bladder cancer cell lines in vitro to determine their reactivity and cytotoxicity. Among these results are GI50 concentrations for each drug, which is the concentration of drug needed to see growth inhibition in 50% of treated cells. Previous published …

Cloning The Vision Related G Protein Transducin For Live Cell Fluorescence Studies, Deanna M. Bowman

Williams Honors College, Honors Research Projects

G coupled protein receptors (GCPR) are one of the largest families of receptors and mediate a variety of biological responses. Rhodopsin is the largest family and aids in sight, the α-subunit of the GCPR complex in extremely important to the activation and downstream signaling effects of GCPR. The α-subunit contains a small trans-domain portion and in this project the sequence of that portion will be inserted into a vector containing a fluorescent tag. These vectors will then be used to collect fluorescent cross correlation spectroscopy or FCCS data. The unit was cloned using assembly methods that include PCR and purification …

Effect Of Osteoactivin And Bone Morphogenetic Protein-2 On Soft Tissue In Rat Spinal Fusion Model, Andrew Ohliger

Williams Honors College, Honors Research Projects

Osteoinductive therapeutic strategies are essential for the bone healing process and are applicable in multiple orthopaedic procedures. The novel protein Osteoactivin (OA) has proven to stimulate osteoblast differentiation and function in vitro and bone regeneration in vivo in the rat model. More recently, OA has shown significant bone regeneration ability in sheep. The goal of this study is the comparison of osteoactivin with bone morphogenetic protein-2 (BMP-2) in the rat spinal fusion model.

Investigating Iron Metabolism In Subarachnoid Hemorrhage Patients, Alexander A. Maynard, Gardenia Pacheco

Williams Honors College, Honors Research Projects

Subarachnoid hemorrhage (SAH) is a stroke characterized by bleeding into the subarachnoid space of the brain, typically resulting in high mortality rate.⁸ Delayed cerebral ischemia (DCI), characterized by vasospasms induced arterial constriction, occurs in roughly one third of the surviving patients.²⁰ The development of DCI and neurodegeneration could be linked to metabolic pathology that occurs after SAH, specifically iron induced changes in redox status. The oxidized environment induced by iron has the potential to functionally affect the ferroxidase ceruloplasmin (Cp), which is linked to neurodegeneration.¹⁵ Global LC-MS based metabolomics data revealed alterations in metabolism in the CSF …

Detection Of Semi-Polar Vitamins And Nutrients In Biological Samples, Jennifer Janovick

Williams Honors College, Honors Research Projects

Metabolomics is used to examine metabolite fluctuations in biological systems and enables the diagnosis of metabolic disorders and the identification of new therapeutic targets.¹ Liquid chromatography-mass spectrometry (LC-MS) was utilized to develop a new method using a T3 column to separate small molecule nutrients and vitamins in a tissue sample. Initial tests of column performance used standard solutions; the amount of metabolites identified and the elution profile of these metabolites was examined. Both the standard solution and sample of brain tissue were tested with the final method developed. Six metabolites were identified in the standard and 31 were identified …

Mutations In Rhodopsin Transmembrane Helix 5 And Dimerization Interface, Margaret Pinkevitch

Williams Honors College, Honors Research Projects

G protein coupled receptors (GPCRs) are membrane proteins with many functional roles. This family of receptors is expansive, with several subfamilies. Here, a member of subfamily A, rhodopsin (Rho) is studied. Mutations in rhodopsin have been linked to the disease retinitis pigmentosa (RP). Some of these mutations have been found to disrupt the dimerization/oligomerization of Rho at the TM4/TM5 proposed dimerization interface, causing pathogenesis of RP. In this study, two mutations of transmembrane helix 5 were investigated for changes in dimerization and stability, cell trafficking, and signaling. It was found that the V209M and F220C mutations did not result in …

Sporadic Appearance Of Paralytic Spongiform Neurodegeneration In A Colony Of Inbred Mice Is Associated With Cns Retrovirus Expression, Magdalene Durbak

Williams Honors College, Honors Research Projects

Abstract

Exogenous and endogenous retroviruses (RV) have been known to induce vacuolar central nervous system neuropathology. Two mice which were juvenile inbred Rocky Mountain White (IRW), showed signs of spontaneous degenerate motor neuron disease. Signs included wasting, unkempt fur, adduction reflex upon tail elevation, kyphosis, and stilted gait by postnatal day 18 (P18). Brains and spinal cords were examined with H&E staining and stained with a broadly reactive retrovirus antibody. When compared to brains of mice with normal vacuolar pathology as well as mice infected with prototypic non-neurovirulent RV, Fr57E and prototypic neurovirulent RV FrCasE, the subject showed less severity …

Altering Oligomerization Of Epha2 Via Mutations In The Intracellular Domain, Ryan W. Lingerak

Williams Honors College, Honors Research Projects

Eph receptor tyrosine kinases (RTKs) are activated by membrane-bound ligands called ephrins. Eph RTKs are divided into two subclasses, each activated by a specific classes of the ligand ephrin. The overexpression of Eph receptors is correlated to cancer cell metastasis in several different types of cancers. Studies with the EphA2 extracellular domain (ECD) and ephrinA1 ligand have shown that upon binding of ephrin to the receptor, EphA2 undergoes increased oligomerization and activation. This indicates that oligomerization is intimately connected to kinase activity. High resolution crystal structures of the EphA2 ECD have revealed some details of these ligand bound oligomers, as …

Effects Of Novel Fragment-Warhead Adducts In Situ And In Vitro With Glutaredoxin Orthologs., Andrew J. Caras

Williams Honors College, Honors Research Projects

Fragment-based drug design (FBDD) is a method of pharmaceutical research involving characterization of small “fragment” molecule interactions with a target protein. These fragments are optimized with the addition of a nucleophilic “warhead” moiety. Several novel fragment-warhead adducts have been synthesized in house by group member Ram Khattri Ph.D. The experiment investigates reactivity of these adducts with glutaredoxin protein (Grx) orthologs from Homo sapiens, Pseudomonas aeruginosa and Brucella melitensis. Three novel fragment-warhead adducts have been identified to cause weakly bacteriostatic and bacteriocidal effects, most with minimum inhibitory concentration values near 1.28 mg/mL. Discussed herein are molecular effects of these fragment-warhead …

Analysis Of Microbial Diversity In Disturbed Soil, Tyler G. Sanda

Williams Honors College, Honors Research Projects

This paper uses the composition and abundance of microbial species to analyze soil recovery in disturbed land. Surface mining disturbs ecological communities throughout the world. As organizations seek to reclaim these disturbed lands, a proper analysis of recovery is needed. In previous studies, recovery of disturbed land was limited to surface examinations, which do not characterize the possible unseen devastating effects of the subsoil. Soil microorganisms are extremely sensitive to environmental changes such as strip mining. It is proposed that these microorganisms may serve as better indicators of recovery post disturbance. Our analysis indicates microbial recovery, however it may not …

Nanoparticle Targeting To The Central Nervous System, Emily Baker

Williams Honors College, Honors Research Projects

Drug delivery to the central nervous system is complicated by the blood-brain barrier, a vascular structure that prevents free diffusion of molecules into brain tissue. In this study, we examined the in vitro properties of a nanoparticle-based drug delivery system. We used several nanoparticle formulations, both LTP based and PLGA based, to test the cellular uptake and toxicity in microglial cells. Using immunofluorescence imaging, we show that LTP nanoparticles are taken up by microglia. We confirmed that our nanoparticle formulations are non-toxic by two cell viability assays. These results suggest that nanoparticle formulations may be a biocompatible method of delivering …

The Role Of Tgfβ1 And Ep Receptors In The Differentiation Of Normal And Diseased Lung Fibroblasts, Madison A. Hexter

Williams Honors College, Honors Research Projects

Fibroblasts, and their differentiation into myofibroblasts, are important in the pathology of several lung diseases including asthma and idiopathic pulmonary fibrosis (IPF). The differentiation process is dependent on several factors, such as soluble factors like transforming growth factor beta 1 (TGFβ1) and mechanical signals such as matrix stiffness. Additionally, the soluble arachidonic acid metabolite Prostaglandin E₂ (PGE₂) prevents TGFβ1 induced differentiation of fibroblasts to myofibroblasts, but the molecular mechanism is not completely understood¹⁵. Transient Receptor Potential Vanilloid 4 (TRPV4), a mechanosensitive ion channel, has previously been implicated in the differentiation of cardiac fibroblasts^12,13. …

Lipid-Protein Interactions Via Fluorescence Quenching In Small Unilamellar Vesicles, Talia Kordahi

Williams Honors College, Honors Research Projects

Studying peptide interactions with model membranes have aided in understanding protein-lipid interactions more clearly. In this report, fluorescent small unilamellar vesicles (SUVs) of a known composition were made. Two peptides, RND1 and MARCKS, were also fluorescently labeled and added to a solution of SUVs. The interactions between these were observed via fluorescence quenching, read out with a spectrofluorimeter. In addition, two environments of buffer were tested: 1X PBS and 0.1X PBS. After obtaining data, it was concluded that MARCKS quenched the SUVs more than RND1. This could be due to the presence of more lysine amino acids in MARCKS, which …

Investigation Of Dna Interaction With Bis-Cationic Imidazolium Salts, Kyle Daily 6440928

Williams Honors College, Honors Research Projects

Imidazolium salt compounds have shown promise in vitro against non-small cell lung cancer (NSCLC) lines, but the exact mechanism of activity these compounds is still unknown. Due to their planar aromatic structure, we hypothesized that our bis-cation imidazolium salt series could act through DNA intercalation similar to known intercalators ethidium bromide, acridine orange, and echinomycin. Here, we tested bis-cation imidazolium salts with linkers of 2, 4, 10, and 12 carbons for DNA intercalation via viscosity and fluorescent intercalator displacement (FID) assays. Despite some solubility issues that may have decreased results, we found that our butyl and dodecyl linked compounds had …

Evaluation Of The Signature Molecular Descriptor With Blosum62 And An All-Atom Description For Use In Sequence Alignment Of Proteins, Lindsay M. Aichinger

Williams Honors College, Honors Research Projects

This Honors Project focused on a few aspects of this topic. The second is comparing the molecular signature kernels to three of the BLOSUM matrices (30, 62, and 90) to test the accuracy of the mathematical model. The kernel matrix was manipulated in order to improve the relationship by focusing on side groups and also by changing how the structure was represented in the matrix by increasing the initial height distance from the central atom (Height 1 and Height 2 included).

There were multiple design constraints for this project. The first was the comparison with the BLOSUM matrices (30, 62, …

Sharp Rna Recognition Motif Optimizations, Extensions, And Mutations For Use In 2d And 3d Nmr Experiments, Shaun M. Christie

Williams Honors College, Honors Research Projects

SMRT/HDAC Associated Repressor Protein interacts with the long noncoding RNA, produced by SRA, by binding at the RRMs. Three projects were done to prepare the truncated proteins for use in 2D and 3D NMR experiments. The first focuses on RRM 3 and its optimization during the purification process. The second focuses on RRM 2-4, which was found to be missing two alpha helices that may be important for protein stability. These helices can also interact with RRM 3 as well due to the tight association of RRMs 3 and 4. The two step PCR extension of RRM 2-4 was assumed …