Biochemistry, Biophysics, and Structural Biology Commons^™

Characterization Of 3d Printed Microdialysis Probes For Solute Recovery And Non-Specific Adsorption, Bailey N. Scoggins

Chemistry & Biochemistry Undergraduate Honors Theses

Microdialysis is an important sampling technique that has uses in a wide range of disciplines, such as neuroscience and pharmacokinetics. 3D printing aims to address adaptability and customization issues that are present in standard “off-the-shelf” microdialysis probes. This work aims to lay the groundwork for characterizing a 3D printed microdialysis probe designed at the University of Arkansas. One experiment involved measuring solute recovery of caffeine as a function of flow rate of a standard CMA 11 probe to assess its membrane performance. The other goal was to learn about non-specific adsorption tendencies of enzymes (i.e., to what degree proteins stick …

Towards The Design Of A Super Fgf1-Fgf10 Chimera, Megan Brown, Suresh Thallapuranam

Biological Sciences Undergraduate Honors Theses

The Fibroblast Growth Factor family consists of 23 different fibroblast growth factors (FGFs) that are each involved in various cellular processes, including cell proliferation and tissue repair. Fibroblast growth factor receptors (FGFR) are different tyrosine kinase receptors. There are four FGFRs in the human body, each binding to different combinations of FGFs. Fibroblast growth factor 1 (FGF1) is the only FGF that can bind with all four types of fibroblast growth factor receptors. Therefore, it is the most powerful mitogen. However, FGF1 has low thermodynamic stability under the physiological conditions. Prior work in the Kumar lab has constructed several points …

The Expression Of Intestinal Tight Junction Proteins Induced By 1,25-Dihydroxyvitamin D3-Glycosides Supplementation To Reduce Bacterial Chondronecrosis With Osteomyelitis Lameness In Broilers, Kennedy M. Hicks

Biological Sciences Undergraduate Honors Theses

Bacterial Chondronecrosis with Osteomyelitis (BCO) is a leading cause of lameness in broiler chickens, impacting animal welfare and commercial poultry production. Supplementation using organic trace minerals, vitamin D, probiotics, and prebiotics has been studied to mitigate BCO lameness incidences in broiler chickens. Our previous study demonstrated that feeding Panbonis®-G-1,25(OH)₂D₃ from Solanum glaucophyllum, containing 1 µg/kg G-1,25(OH)₂D₃, to broilers for the first 28 days of their production cycle reduced BCO lameness by approximately 50%. In this study, we investigated the impact of 1,25-dihydroxy vitamin D₃ supplementation on the expression of tight junction …

Bpa Adsorption On Microplastics In Different Chemical Conditions, Addison Zehren

University Honors Program Senior Projects

Microplastics (plastic particles < 5 mm in size) are increasingly seen as a health and environmental issue, both for their direct effects when ingested by living organisms and for their ability to adsorb and concentrate other pollutants. Current water treatment processes do not adequately remove microplastics from municipal water supplies, so a thorough understanding of how microplastics affect water quality is needed. The objective of this study is to evaluate the effects of certain water quality parameters on the ability of polystyrene microplastics (PS MPs) to adsorb Bisphenol A (BPA). BPA is an endocrine disruptor (chemically like estrogen) used in plastics which can be released into the environment and act as a water pollutant. Water quality chemical parameters investigated were chloride, nitrate, phosphate, and copper ions. Concentrations of each solute were selected based upon Environmental Protection Agency (EPA) guidelines and previous results obtained from testing Chicago River water samples. A constant amount of PS MPs and BPA were added to each sample and the mixtures were shaken for 5 days to allow time for the BPA to adsorb. The samples were then filtered and the amount of BPA left in the water was determined by measuring its UV absorbance on a UV-vis spectrophotometer. BPA concentrations decreased in the water indicating that it had been adsorbed onto the microplastic surface, with the highest adsorption present with copper, which reduced BPA concentrations from 6.7 mg/L to 3.9 mg/L. Statistical analyses showed differences between copper, phosphate, nitrate, and chloride (df = 9, p = 0.002). Future work can apply these results to investigate the implications of pollutant transportation in urban waterways like the Chicago River.

Fouling-Resistant Voltammetric Xylazine Sensors For Detection Of The Street Drug “Tranq”, Ann Holland Wemple

Honors Theses

As the opioid crisis continues to wreak havoc on a national and global scale, it is increasingly critical to develop methodologies to detect the most dangerous of these drugs as fentanyl and its derivatives have orders of magnitude higher potency than morphine. Recently, the scientific challenge for chemical detection of fentanyl and its derivatives has been complicated by both the constantly increasing synthetic variations of the drug as well as the expanded use of adulterants. One tragically consequential example is the nocuous street drug known as “Tranq” which combines fentanyl or a fentanyl derivative with the veterinary sedative Rompun®, chemically …

Fc Gamma Receptors Facilitate Antigenic Modulation Of Lilrb4 And Function As Predictive Biomarkers In Acute Monocytic Leukemia, Joshua Morse

Dissertations and Theses (Open Access)

Acute monocytic leukemia (monocytic AML) is a subtype of AML marked by a proliferation of abnormal monoblasts. This subtype represents approximately 10% of AML cases. The prognosis of monocytic AML is poor, with a 5-year survival of ~30%. Most patients are diagnosed at an age when they are unlikely to survive first-line non-targeted cytotoxic chemotherapy as a bridge to hematopoietic stem cell transplant (HSCT). Even patients who achieve remission commonly relapse. This population of patients would greatly benefit from precision-targeted therapies but currently there are none approved for monocytic AML.

Leukocyte immunoglobulin-like receptor B4 (LILRB4) is an immune checkpoint expressed …

Expanding Inducible Epithelial Resistance Against Pneumonia: Novel Virus, Novel Receptor, Michael Longmire

Dissertations and Theses (Open Access)

Pneumonia is the leading infectious cause of death worldwide and presents a continuing risk among immunocompromised patients. Our work has been focused for many years on the phenomenon of inducible epithelial resistance, where activation of the innate immune response in the airway epithelium is necessary and sufficient to produce a pathogen-killing but self-protective environment to prevent severe disease. Inducible resistance is pathogen-agnostic and has been demonstrated against an array of respiratory infections. We achieve this using a combination of pattern recognition receptor agonists – Pam2CSK4 and ODN M362 being the current formulation – which produce synergistic levels of pathogen reduction …

Development And Functional Characterization Of Carbon Nanodot-Peptide Conjugates For The Treatment Of Bone Disease, Abigail Ali

Honors Theses

Carbon nanodots (CDs) synthesized from carbon nanopowder under acid oxidative conditions have unique bone-bind properties that could be exploited for theranostic purposes. In the zebrafish vertebrate model system, CDs bind to bone with high affinity and specificity, they don’t appear to bind to other tissues and they do not affect the bone physiology or animal viability. Thus, carbon-derived CDs could be developed for the delivery of pharmacological agents to bones to treat diseases such as osteoporosis. Biologicals such as peptides, proteins and antibodies are a new and exciting class of pharmacological agents that have been successfully used in treating complex …

Isolation And Identification Of Active Antimicrobial Compounds In Alcaligenes And Chromobacterium Strains, Hannah Lwin

Honors Theses

ESKAPE pathogens comprise six bacteria that can be highly virulent and are likely to develop antibiotic resistance to many of the currently available antibiotics. New antibiotics are needed to combat these pathogens. Our approach was to isolate and identify antimicrobial compounds from bacteria isolated from soil. Two isolates demonstrating antimicrobial activity were identified as Chromobacterium sp. and Alcaligenes parafaecalis. Preliminary extraction of their antimicrobials was performed.

Effect Of Glycation On The Binding Interactions Of Serum Albumin And Silver Nanoparticles, Megan Kirk

Celebrating Scholarship and Creativity Day (2018-)

Nanoparticles are particles ranging from 1-100 nanometers in size. Most nanoparticles, upon contact with biological matrices, are immediately coated with proteins, forming the protein corona, which can alter the structure and function of proteins. Despite the tremendous advancement of nanotechnology over the last decade, the interactions of nanoparticles with proteins are not well characterized. This project investigates the effect of glycation on the binding interactions of HSA, the most abundant protein in human blood, with silver nanoparticles using various spectroscopic techniques. This is important because in diabetic patients, HSA is exposed to high concentrations of glucose over extended periods of …

Implications Of Type Iv Pilus Retraction For Dna Uptake By Acinetobacter Baumannii, Yafan Yu

Dissertations and Doctoral Documents, University of Nebraska-Lincoln, 2023–

Horizontal gene transfer (HGT) via natural competence allows bacteria to incorporate extracellular DNA (eDNA) into their genomes, facilitating genetic diversity and adaptation. In Acinetobacter species, natural competence depends on DNA binding and uptake mediated by type IV pili (T4P). T4P are dynamic extracellular filaments composed primarily of the major pilin subunit PilA, along with a few minor subunits. T4P drive a range of cellular functions including twitching motility, biofilm formation, and DNA uptake, with some functions dependent on pilus retraction and others not. However, how Acinetobacter T4P bind DNA and how T4P/eDNA interactions impact biofilm formation remain unclear. Here, I …

Screening For Chloroplast Inner Envelope-Thylakoid Membrane Contact Site Proteins, Evan W. Labrant

Dissertations and Doctoral Documents, University of Nebraska-Lincoln, 2023–

Most life on Earth is dependent upon the capture of solar energy and subsequent fixation of carbon. In members of the Archaeplastida lineage these biological processes take place in semi-autonomous organelles called chloroplasts. Chloroplasts are bounded by a double membrane envelope and contain the photosynthetic membrane system, thylakoid membranes, where light capture takes place. While a great deal is known about how thylakoid lipids are synthesized and protein complexes therein are properly maintained, very little is known regarding the processes by which lipids are transported to thylakoids which lack de novo glycerolipid synthesis. We have worked to identify candidate proteins …

Treating The Black Fever: Evaluating Combination Chemotherapy For Leishmania Donovani, Robert Frank Mickatavage

Biochemistry and Molecular Biology

Visceral leishmaniasis (VL) is a neglected tropical disease (NTD) endemic to Africa, Asia, and the Americas, caused by the parasite L. donovani and spread by the Phlebotomine sandfly. If left untreated, VL is almost always fatal, bearing a mortality rate of over 95%. Many individuals within endemic regions are often too poor to afford treatment, and the financial burden of current treatments may only exacerbate their poverty. Antimonials and more modern treatments display varying effectiveness, but are nonetheless toxic to the host, commonly inducing hepatotoxicity, nephrotoxicity, and renal failure. The development of more effective and safer treatments is crucial, particularly …

Sodium And Potassium In Health And Cellular Function, Jordyn Branz

Honors Program: Senior Projects (Public)

Sodium (Na⁺) and potassium (K⁺) are essential electrolytes that regulate numerous physiological processes. These include cell function, nerve transmission, and muscle contraction. The Na+/K+-ATPase plays a critical role in maintaining the balance of these ions by actively transporting three sodium ions out of the cell and two potassium ions into the cell against their concentration gradients. This process is vital for maintaining the resting membrane potential and supporting the function of excitable cells. The Na+/K+-ATPase consists of two main subunits, with the alpha-subunit being primarily responsible for ion transport activity. Isoforms alpha1, alpha2, and alpha3 are tissue-specific and contribute to …

Implications Of Acetylation And Phosphorylation On Metabolic Enzymes, Nour Fatema

Graduate Theses and Dissertations

All living things depend on glycolysis and the TCA cycle for energy production and metabolic control. Post-translational modifications (PTMs) such as acetylation and phosphorylation, which can change enzyme activity, substrate binding, and overall metabolic balance, tightly regulate these pathways. Diseases like cancer and metabolic problems are associated with disruptions in these alterations.

Serine phosphorylation and lysine acetylation are two of the many PTMs that are evolutionarily conserved processes that alter protein function. Previously thought to be a static histone alteration, acetylation is now understood to be a dynamic regulator of metabolic enzymes that modifies contacts, charge, and conformation. In response …

Intratumoral Fibrin As A Novel Immunomodulatory Factor In Pancreatic Ductal Adenocarcinoma, Mazharul Karim

Open Access Theses & Dissertations

In-situ coagulation is a common clinical feature of pancreatic ductal adenocarcinoma (PDAC) that presents a highly thrombotic and crosslinked insoluble fibrin (x-fibrin)-laden tumor stroma (FibTS). However, the specific impact of FibTS on immune cells behavior remains largely unexplored in this poorly infiltrated tumor. We aimed to elucidate how FibTS in PDAC regulates immune cell infiltration, polarization, and crosstalk that favors immunosuppressive microenvironment and tumor growth. We characterized human PDAC tissues by multiplex immunostaining and checked the spatial distribution of immune cells based on the presence of x-fibrin. We investigated how FibTS influences the infiltration of tumor-associated macrophage (TAM) and T-cell …

Jak1 V658f Drives Oncogenic Transformation Via Transcriptional Reprogramming Independent Of Y598 Phosphorylation And Canonical Jak-Stat Signaling, Omar Javier Rodriguez Moncivais

Open Access Theses & Dissertations

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive and complex malignancy that accounts for 15% of pediatric acute lymphoblastic leukemia (ALL) cases in the United States. Tyrosine kinases control critical cellular processes, including the development, survival, and activation of T cells, and are established drivers of leukemia.Tyrosine kinases involved in hematological disorders include receptor tyrosine kinases and non-receptor tyrosine kinases family members such as SRC, ABL, FAK, and Janus kinases (JAKs). This study primarily focused on JAK1 gain-of-function (GoF) mutations. These mutations are recognized as hyperactivating mutations, maintaining kinase activity of JAKs on and essentially keeping downstream signaling effectors, like …

A Qualitative Nmr And Histological Analysis Of Adipocere Formation In Different Aqueous Environments Using Sus Scrofa Models, Mia M. Sorrentino

Student Theses

Adipocere is a wax-like crust that has the potential to form over corpses in moist, humid, and anaerobic environments, preserving them in a process similar to that of mummification. The substance can be identified within 3-6 months postmortem and is characterized by the ratio of saturated fatty acids to unsaturated fatty acids. Palmitic acid and stearic acid are the most abundant in comparison to myristic and oleic acids. The goal of this study was to investigate factors affecting adipocere formation in different aqueous environments. Pork belly samples were submerged in water samples collected from different places around New York City …

Developing A Small Molecule To Inhibit Hsf1 Expression In Cancer And Evaluating Natural Genetic Variation In Small Molecule Toxicity., Michaela Kendal Foley

Theses and Dissertations

Each year cancer affects nearly 20 million people worldwide and genetic differences across populations can impact cancer onset and progression. Specifically, tumors with high levels of HSF1, the master regulator of the cytoprotective heat shock response (HSR), are correlated with poor patient outcomes in multiple cancers such as prostate, breast, and melanoma. Subsequently, the development of pharmacological inhibitors of HSF1 represents a promising strategy for anticancer therapeutics. Using a luciferase-based transcriptional reporter, two small molecule libraries were screened for inhibitors of HSF1 expression in human embryonic kidney cells, yielding ten compounds that decrease HSF1 expression. To identify if cancer lines …

Identification Of Cadmium As A Structural And Functional Modulator Of Psd-95 Regulating Nmdars At Postsynaptic Membranes, Claudia Garza

Theses and Dissertations

Postsynaptic density-95 (PSD-95) is an essential scaffolding protein in excitatory neurons. PSD-95’s N-terminal palmitoylation region elicits synaptic plasticity by targeting, anchoring, and trafficking glutamate receptors, such as N-methyl-d-aspartate receptors (NMDAR), to the postsynaptic membrane. The molecular interactions of PSD-95 and its dysregulation in excitatory synapses are currently being explored as primary causes of neurodegenerative diseases and disorders, such as Alzheimer’s disease, Schizophrenia, and Parkinson’s disease. Recent studies demonstrate an overaccumulation of zinc at two cysteines (C3 and C5) and two histidines (H24 and H28) of PSD-95 blocks palmitoylation, causing a loss of NMDAR at the postsynaptic membrane. In addition, exposure …

Cloning And Heterologous Expression Of Human Nachrs And A Unique Spider Venom Toxin, Christine M. Vega

Theses and Dissertations

Drug development research has long focused on synthesizing novel therapeutics while overlooking isolation from naturally available sources. Recently, the discovery of the therapeutic effects of highly bioavailable animal venom caused the field of venom research to soar in popularity. Animal venom serves as a natural, sophisticated tool optimized over millions of years by evolution with the ability to bind to ion channels such as nAChRs, non-specific ligand-gated ion channels distributed throughout the human nervous system, with high selectivity and specificity. However, purifying individual venom toxins from crude venom is challenging due to its availability. Therefore, synthesizing large quantities of venom …

Effects Of Cholesterol Concentration On Liposome And Proteoliposome Behavior, Ehsaneh Khodadadi

Graduate Theses and Dissertations

Cholesterol is crucial in modulating the behavior and characteristics of biological membranes, liposomes, and proteoliposomes, for instance in drug delivery applications. Cholesterol regulates membrane stability by influencing lipid density, phase separation, and protein-membrane interactions. In planar bilayers, cholesterol enhances membrane thickness, reduces permeability, and promotes lipid ordering, adding rigidity and fluidity. Cholesterol also modulates membrane curvature in curved bilayers and vesicles, stabilizing low-curvature regions. In this work, all-atom molecular dynamics (MD) simulations were employed to examine how varying cholesterol concentrations influence the structure and dynamics of membranes, focusing on planar and curved geometries. Bilayers with cholesterol molar ratios of 0%, …

Bang!: Real-Time Measurements And Transcriptomics Of Metabolic Reprogramming In Activated Macrophages At Fever Temperature, Kenneth Howard Taylor

Graduate Theses and Dissertations

We investigated the real-time metabolic rates of activated macrophage cell lines derived from mouse, chicken and rainbow trout. Each species showed a different kinetic signature with metabolic rates accelerating under fever conditions specific to each organism. Upon activation mouse macrophages increased their rate of glycolysis early and then a later reprogramming phase occurred with a dramatic decrease in oxidative phosphorylation (OXPHOS) accompanied by a reciprocal second increase in glycolytic rates. Chicken macrophages showed an initial increase in both glycolytic and OXPHOS rates. The late drop OXPHOS rates also occurred in chicken however there was no reprogramming that occurred as glycolytic …

The Regulatory Role Of Collagen Α1(Xi) Amino-Terminal Domain Isoforms In Collagen Fibrillogenesis, Abu Sayeed Chowdhury

Boise State University Theses and Dissertations

Collagen α1(XI) is a minor fibrillar collagen involved in the critical regulation of collagen fibrils, such as nucleation, assembly, and regulation of fibril diameter. The amino-terminal domain consists of an amino-propeptide (Npp) domain and a variable region, which is the result of alternative splicing from exon 6A, 6B, 7, and 9. The spliced variants involving Npp, with the combinations of exon 6A, 6B, 7, and 9 gives rise to different isoforms with very distinctive chemical nature. The Npp of collagen α1(XI) chain is retained on the surface of the collagen fibril for an extended period of time and may play …

Impact Of Tumor Cell Expressed Cd38 On Metastasis And Immune Evasion In Breast Cancer, Tanvi Visal

Dissertations and Theses (Open Access)

Triple-negative breast cancer (TNBC) is a highly metastatic breast cancer subtype. The epithelial-to-mesenchymal transition (EMT) of cancer cells is a key feature of the metastatic cascade and is not a binary process but often generates malignant cells with both epithelial (E) and mesenchymal (M) traits known as hybrid EM cells. Recent studies highlight the enhanced metastatic potential of the hybrid EM cells. However, molecular insights and targetable vulnerabilities within hybrid EM remain elusive. We discovered that hybrid EM murine tumors are enriched in CD38, an immunesuppressive molecule associated with worse clinical outcomes in liquid malignancies but relatively understudied in solid …

The Extracellular Domains Of The Ampa, Kainate, And Orphan Delta Receptor Regulate Channel Gating In Distinct Ways, Cuauhtemoc Ulises Gonzalez

Dissertations and Theses (Open Access)

The ionotropic glutamate receptors are ligand-gated cationic channels essential for neuroplasticity and neurological functions such as learning, memory, behavior, cognition, and motor coordination. Mutations in these channels are linked to various disorders including autism, ataxia, and epilepsy, making them compelling pharmacological targets. Within this family, the α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) and kainate receptor mediate their function through fast glutamate-gated ionotropic activity, which needs precise structural changes to perform properly. While Cryo-EM structures have provided snap shots of these proteins, questions remain regarding the dynamics across conformations and individual roles of the extracellular amino-terminal and agonist binding domain in dictating function. …

Overcoming Resistance To Cdk4/6-Targeted Therapy Using Jak2/Stat3 Inhibitor In Triple-Negative Breast Cancer, Chuling Zhuang

Dissertations and Theses (Open Access)

Breast cancer is the most common type of cancer diagnosed in women, with nearly 30% of cases becoming metastatic accounting for over 90% of breast cancer-related deaths. Among different breast cancer subtypes, triple-negative breast cancer (TNBC) has the worst prognosis and the highest propensity to metastasize. However, TNBC patients have limited treatment options due to their lack of actionable drug targets while therapeutic resistances result in high rates of recurrence and metastasis. Cyclin-dependent kinase 4 and 6 (CDK4/6) are major cell cycle regulators that control G1 to S phase transition and aberrant hyperactivation of the CDK4/6 pathway results in uncontrolled …

Role Of Mir-4732-3p In Breast Cancer Brain Metastasis And Brain Metastatic Tumor Microenvironment, Munazza Samar Khan

Dissertations and Theses (Open Access)

In this study, we aimed to identify microRNAs (miRNAs) that may play important roles in breast cancer brain metastasis (BCBM). To this end, we conducted miRNA-sequencing of extracellular vesicles isolated from the serum samples of 6 BCBM patients and 8 Stage I/II/III breast cancer patients, and identified 49 circulating miRNAs that were upregulated in BCBM patients compared to Stage I/II/III breast cancer. Upon further analysis, we identified miR-4732-3p to be upregulated in brain-tropic triple-negative breast cancer (TNBC) cell lines, compared to parental lines. MiR-4732-3p overexpression increased metastatic properties of TNBC cells, including proliferation, migration, invasion, and maintenance of a mesenchymal …

Investigating Metabolic And Mitochondrial Adaptations Triggered By Blood Flow During The Endothelial-To-Hematopoietic Transition, Paulina Horton

Dissertations and Theses (Open Access)

Hematopoietic stem cell (HSC) transplant is the standard of care for many hematologic diseases. However, many patients cannot benefit from this potentially curative treatment because they cannot find a suitable donor, contributing to a high level of unmet need. Recently, bona fide HSCs capable of robust engraftment and differentiation have been generated from iPSCs utilizing wall shear stress (WSS) to help promote HSC generation. However, the mechanisms through which WSS regulates hematopoiesis remain poorly understood. Scientists, therefore, continue to look for the mechanotransduction pathways that promote hematopoiesis, and key answers may lie in examining the role of extrinsic factors that …

Leveraging Natural Variation In Yeast To Understand Susceptibility Differences In Parkinson’S Disease, Julio Andres Molina Pineda

Graduate Theses and Dissertations

Parkinson’s Disease (PD) is a neurodegenerative disorder that causes countless suffering around the world. Both forms, familial and sporadic, have been associated with the misfolding and cytotoxicity of α-synuclein, an otherwise non-pathogenic protein suspected to have various roles in the normal maintenance of the central nervous system. Model organisms have emerged as great vessels to uncover the cellular and molecular mechanisms that shape α-syn biology, and great strides have been made to understand it. Yet, it is still unclear why some individuals are affected by α-syn toxicity, while others not so much. Since it’s extremely complicated to study and probe …