Biochemistry, Biophysics, and Structural Biology Commons^™

Role Of Grb2 Intracellular Localization In Progesterone Receptor Positive Breast Cancer, Nattamolphan Wittayavimol

Chulalongkorn University Theses and Dissertations (Chula ETD)

Progesterone receptor (PR) extranuclear signaling mediates mainly through PR polyproline domain (PPD) interaction with SH3 domain of cytoplasmic signaling molecules. PR-PPD interfered with Grb2-SOS binding through SH3 interaction resulting in decreased cell proliferation and correlates with a good prognosis in PR positive breast cancer. Grb2 is a key adaptor molecule and expressed in both nucleus and cytoplasm. We hypothesized that change in Grb2 localization alters PR transcription resulting in better clinical outcomes in PR positive breast cancer. BT-474 breast cancer cells expressing inducible Grb2-SV402x (nuclear Grb2, nuGrb2) or Grb2-MYR (membrane Grb2, memGrb2) were constructed. MemGrb2-BT-474 showed higher levels of PR …

Roles Of Progesterone Signaling On Dna Repair And Cell Survival In Ovarian Cancer Cells, Pattarasiri Rangsrikitphoti

Chulalongkorn University Theses and Dissertations (Chula ETD)

Every year over 300,000 women worldwide are diagnosed with ovarian cancer and half of these patients die from this disease. The symptoms of ovarian cancer are very general and often confused with other common female diseases making ovarian cancer a challenging disease to diagnose and treat. Platinum-based chemotherapy, including cisplatin, stands as the first-line treatment for ovarian cancer. However, chemotherapy resistance remains a continuing problem, contributing to a high mortality rate. Progesterone receptor (PR) signaling has been implicated in various aspects of ovarian cancer biology. Clinical studies show a positive correlation between PR expression, improved patient survival and cisplatin response, …

Cisplatin - Loaded Calcium Citrate Nanoparticlesfor Inhibition Of Lung Cancer Cell Growthand Quantum Mechanical Study Of Cisplatin And Citrate, Lipika Oopkaew

Chulalongkorn University Theses and Dissertations (Chula ETD)

The International Organization for Research on Cancer (International Agency for Research on Cancer (IARC) issued a statement saying lung cancer is a cancer with high incidence rates of new cases and deaths. Although there are current therapeutic approaches such as surgery, radiation therapy, and chemotherapy. However, these treatments exhibit ineffectiveness; the main limitation is severe side effects to normal cells or non-target organs. To address this challenge, the development of targeted therapy by nanoparticle-based drug delivery system is applied to increase drug stability and control the specificity to cancer cells with target-specific proteins that play an important role in intracellular …

Protein Extraction And Purification By Differential Solubilization, Barry J. Ryan, Gemma K. Kinsella, Gary T. Henehan

Books/Book Chapters/ Proceedings

The preparation of purified soluble proteins for biochemical studies is essential and the solubility of a protein of interest in various media is central to this process. Selectively altering the solubility of a protein is a rapid and economical step in protein purification and is based on exploiting the inherent physicochemical properties of a polypeptide. Precipitation of proteins, released from cells upon lysis, is often used to concentrate a protein of interest before further purification steps (e.g., ion exchange chromatography, size exclusion chromatography etc).

Recombinant proteins may be expressed in host cells as insoluble inclusion bodies due to various influences …

Apobec3 Regulation By Thyroid Hormone, Michelle Santiago

All Graduate Theses, Dissertations, and Other Capstone Projects

Thyroid hormone (TH) is crucial for many biological processes, including the development of the auditory system. Appropriate thyroid hormone signaling is important for the development of the cochlea, leading to the successful onset of hearing. More specifically, TH influences the timing of remodeling of the greater epithelial ridge (GER), a transient cochlear structure that secretes proteins that contribute to the formation of the tectorial membrane. Developmental hypothyroidism results in delayed GER regression, whereas excess TH results in premature remodeling. Both conditions result in deafness. However, it is unknown how TH mediates GER remodeling. Expression profiling identified APOBEC3 (A3) as a …

The Molecular Mechanisms Of Ifn-Γ Promoted Necroptosis In Macrophages, Meghan R. Vogt

Biotechnology Theses

Necroptosis, a form of programmed lytic cell death, has emerged as a driving factor in the pathogenesis of acute lung injury. One specific inflammatory mediator that is of interest is interferon-γ (IFN-γ). Our laboratory has reported that IFN-γ is the most potent enhancer of necroptosis in primary lung epithelial cells and macrophages by upregulating MLKL expression. How IFN-γ enhances necroptosis and MLKL upregulation is currently unknown. In this research, we utilized multiple approaches to study the potential molecular mechanisms of IFN-γ promoted necroptosis in macrophages. We found that MLKL and ZBP1 are significantly upregulated by IFN-γ through an unknown pathway …

Investigating The Relationship Between Metabolic Reprogramming And Peripheral Cd4+ T-Cell Inflammation In Human Type 2 Diabetes Pathogenesis, Gabriella Kalantar

Theses and Dissertations--Microbiology, Immunology, and Molecular Genetics

Chronic, low-grade systemic inflammation rises in obesity and promotes type 2 diabetes (T2D). Circulating immune cells are key indicators of obesity and T2D pathogenesis. T cells outnumber monocytes, in blood, suggesting that T cells might fuel peripheral inflammation in obesity/T2D. Our lab’s work supports this idea by identification of a Th17 cytokine profile in T2D from T-cell stimulated peripheral blood mononuclear cells. Work described herein further supported this work by demonstrating that T cells dominate peripheral inflammation over monocytes across the spectrum of obesity and glycemic control. Our lab has also recently shown that inflammation changes during prediabetes (preT2D), identified …

Developing A Biocatalytic Toolbox To Aid In Understanding Nucleoside Antibiotics, Jasmine Brianna Woods

Theses and Dissertations--Pharmacy

Antibiotic resistance happens when bacteria develop the ability to survive medications that normally terminate them. Instead, these super germs are able to survive in the body and produce a community of antibiotic resistance germs which can cause human fatalities. It is important to discover and develop new compounds and molecules that will improve this clinical obstacle. This research focused on analyzing the biosynthesis that incorporates distinctive chemical characteristic of various nucleoside antibiotics, ß-hydroxy amino acids and α-methyl-amino acids. ß-hydroxy amino acids and α-methyl-amino acids are considered an important class of industrially useful compounds, particularly for pharmaceutical development, and are found …

Impact Of Sample Conditions On Dna Phosphodiester Backbone Bi/Bii Conformational Equilibrium Dynamics, Autumn C. Pilarski

Graduate Theses/Dissertations

DNA damage, such as single base lesions and mismatches, is highly prevalent within cells. If these DNA damage events are not repaired, they could lead to mutations and thus disease and cancer. Intricate repair mechanisms are in place to fix these damage events, one such being Base Excision Repair (BER) and associated enzyme: Thymine DNA Glycosylase (TDG). The first step of this repair process, recognition of the lesion by TDG, is not well understood. The following thesis presents results to better understand the fundamental biophysical question of how a DNA lesion within a mismatch context is recognized in a million …

Investigating Mirna Regulation Of The Human Apobec3 Enzymes, William Dietrich

All Graduate Theses, Dissertations, and Other Capstone Projects

The human apolipoprotein B mRNA editing enzyme catalytic polypeptide 3 (APOBEC3, A3) are a family of proteins consisting of seven enzymes, A3A, B, C, D, F, G and H, which function as cytosine deaminases. The enzymes’ purpose in the cell is to mutate viral DNA during infection hindering or stopping replication of viruses such as human papillomavirus, herpes simplex virus, and HIV-1. Several of the A3 enzymes have also been implicated in contributing to cancers such as head and neck and breast cancers by mutating cellular genomic DNA, making the ability to control A3 expression an attractive target for cancer …

Novel Amino Acids Dictate Differences In Sivmac239 And Hiv-2 Rod9 Vpx Cellular Localization, Hunter D. Mayhew

All Graduate Theses, Dissertations, and Other Capstone Projects

Vpx is a protein encoded by Human Immunodeficiency Virus Type 2 (HIV-2) and specific Simian Immunodeficiency Viruses (SIVs) that is essential for successful infection of non-dividing myeloid cells. Vpx facilitates viral replication by trafficking the viral genome into the nucleus of infected cells and targeting the primarily nuclear innate immune protein SAMHD1 for proteasomal degradation. Considering that these activities occur at the nucleus, we hypothesized that Vpx nuclear localization would be tightly correlated with these functions. To assess this, site directed mutagenesis in combination with confocal microscopy or Western Blotting analyses were utilized to evaluate the determinants of nuclear accumulation …

Investigation Of Mechanical Regulation On Stat3 Activity And Mmp Production, Jaxson R. Libby

Honors Theses and Capstones

Transcription factor, STAT3, is inappropriately expressed in cancer cells, and has contrasting activation in 2D versus 3D microenvironments. 2D plates are often used for drug screening and do not always recapitulate in vivo responses. To combat inaccurate 2D drug studies, a 3D hydrogel was created to support the growth of cancer cells into a tumor-like environment. The hydrogel consists of a biocompatible dextran homopolysaccharide, cell adhesion RGD sequences, and crosslinker MMP labile peptides. A pH dependent reaction couples the RGD sequences to dextran then the polymers are crosslinked into a gel. Crosslinking is accomplished using terminal cysteine peptide sequences, allowing …

Recent Insights On The Conformational Changes, Functionality, And Physiological Properties Of Plant-Based Protein–Polyphenol Conjugates, Mehnaza Manzoor, Zaria Fozonne Ngabou Tchameni, Zuhaib F. Bhat, Amit K. Jaiswal, Sundeep Jaglan

Articles

Plant proteins are promising sources of nutrition with enhanced health benefits due to their interaction with bioactive compounds like polyphenols. These interactions can form protein–polyphenol complexes with diverse properties and applications in the food, nutraceutical, and pharmaceutical industries. To exploit the potential of these conjugates, it is essential to understand how they are formed, characterized, and delivered. Various techniques such as spectroscopy, thermodynamics, and molecular docking simulations can reveal the mechanism and outcomes of protein–polyphenol binding. Moreover, the surface modification of protein–polyphenol conjugation can enhance the pharmacological effects of different drug molecules and enable their targeted delivery to specific tissues …

Molecular Mechanisms Of Wild-Type And Mutant Prion Formation, Daniel J. Walsh

Dartmouth College Ph.D Dissertations

Prion diseases are a class of infectious neurodegenerative disorders which affect humans and many other mammalian species. The causative agent is a unique pathogen known as a prion or PrPSc, a misfolded form of a host-encoded glycoprotein which replicates by templated conformational change. Distinct strains of prions with unique phenotypic and pathologic presentations appear to be encoded by subtle conformational changes within the misfolded protein. While the general manner of prion transmission is known, our detailed understanding of these mechanisms remains incomplete, limiting efforts for the discovery or design of therapeutic treatments for these fatal diseases. In vitro synthesis of …

Interactomics And Targeted Protein Degradation For Kinase Substrate Discovery, Juan C. Mercado Del Valle

Dartmouth College Ph.D Dissertations

Reversible phosphorylation is one of the most important post translational modifications that has allowed us as a species to quickly adapt to changing molecular environments due to external stimulation. This process is only capable through the activity of kinases to carry out the targeting of specific substrates defined by their recognition motif allowing for selective phosphorylation and activation and inactivation of distinct pathways as well as other changes that permit cell survival. By being so important for the maintenance of the cells disruption often leads to worsening of the cells, leading to various diseases like cancer, immunological and neurodegenerative disorders. …

Mitochondrial Division: Synergizing In Mitochondrial Divisome, Ao Liu

Dartmouth College Ph.D Dissertations

Mitochondria are the energy factories of the cell. The dynamic nature of cells demands routine changes in mitochondrial morphology by fusion and division. The dynamin GTPase Drp1 is a central mitochondrial division protein, driving constriction of the outer mitochondrial membrane via oligomerization. At least four regulatory factors control Drp1 activity on the outer mitochondrial membrane (OMM): 1) receptor proteins (Mff, MiD49, MiD51, and Fis1); 2) actin filaments; 3) the mitochondrial phospholipid cardiolipin (CL); and 4) Drp1 post-translational modifications, of which two phosphorylation sites (S579 and S600) are the most well studied. However, the molecular mechanism of how these factors work …

Biogeography Of Special Metabolites Produced By Ant-Associated Bacteria, Katherine Hansen

Scripps Senior Theses

There is an urgent and growing need for novel antibiotics with the growing prevalence of antibiotic-resistant pathogens. Ecologically-guided discovery is a strategy that focuses on how and where specially evolved metabolites such as antibiotics are used in nature. This strategy reveals unique active molecules with potential for human use. An unexplored niche with great potential for antibiotic discovery exists with Southwestern fungus-farming Trachymyrmex ants and their Actinobacteria symbionts. In exchange for nutrients from the ant, the ants’ bacterial symbiont produces antifungal molecules that protect the ants’ fungal cultivar from invasion by pathogenic microorganisms. They also produce antibacterial molecules to protect …

Analysis Of Biologically Effective Dose For Retroactive Yttrium-90 Trans-Arterial Radioembolization Treatment Optimization, Mj Lindsey

CMC Senior Theses

Trans-arterial radioembolization (TARE) is a protracted modality of radiation therapy where radionuclides labeled with Yttrium-90 (⁹⁰Y) are inserted inside a patient's hepatic artery to treat hepatocellular carcinoma (HCC). While TARE has been shown to be a clinically effective and safe treatment, there is little understanding of the radiobiological relationship between absorbed dose and tissue response, and thus there is no dosimetric standard for treatment planning. The Biologically Effective Dose (BED) formalism, derived from the Linear-Quadratic model of radiobiology, is used to weigh the absorbed dose by the time pattern of delivery. BED is a virtual dose that can …

Determination Of The Effects Of Added Methionine On The Toxicity Of Platinum Drugs In Saccharomyces Cerevisiae, Kassidy Owens, Jonathan White

Longwood Senior Thesis Proposal

Platinum (Pt) drugs are used in about half of all anticancer treatments. A common way to study Pt drugs is by using S. cerevisiae (brewer’s yeast) as a model cellular organism. One of the most commonly-used strains of yeast is BY4741, which is auxotrophic for (cannot synthesize) several small-molecule nutrients, including the amino acid methionine. Therefore, methionine must be supplemented in the cell culture medium. Interestingly, it is known that methionine can react with Pt compounds, including Pt drugs. Methionine-supplemented media continues to be used to study Pt drugs in yeast, especially BY4741, yet the potential for methionine to interfere …

Investigating Visinin-Like Protein 1 And Ubiquitinated Visinin-Like Protein 1 As Mild Traumatic Brain Injury Biomarkers, Yueming Wu

Theses and Dissertations--Chemistry

Traumatic brain injury (TBI) continues to be a significant cause of morbidity and mortality worldwide. Despite significant progress in understanding the complex pathophysiology of TBI, the underlying mechanisms remain poorly understood. The primary brain damage is acute and irreversible. However, secondary brain injuries often develop gradually over months to years, creating an opportunity for critical therapeutic interventions. In the past decade, research on TBI biomarkers has seen significant progress. This progress has been driven by the diverse nature of TBI pathologies and the challenges they present for evaluation, management, and prognosis. TBI biomarker proteins resulting from axonal, neuronal, or glial …

College Of Natural Sciences 2022 Year-End Publication, College Of Natural Sciences

College of Natural Sciences Newsletters and Reports

This is the 2022 issue of the annual College of Natural Sciences year-end publication.

Contents:
[Page] 2 Dean's message
[Page] 3 Department highlights
[Page] 4 Overview of Bold & Blue Campaign
[Page] 5 Dr. Edward Hogan recognition & endowment
[Page] 6 Career milestones
[Page] 7 Student travel and research
[Page] 8 $11 million COBRE grant
[Page] 9 Professional Science Masters & Research highlights
[Page]10 Outreach highlights throughout the state
[Page] 11 2022 events recap – join us in 2023!
[Page] 12 Updates on our VR initiative
[Page] 14 Overview of awards and recognitions from 2022

The Effects Of Age On Working Memory Capacity And Neural Function, Jasmin P. Joshi

CMC Senior Theses

Do age-related differences in working memory capacities predict specific neural responses? Theories of working memory (WM) suggest that there are different types of WM processes that are associated with specific neural networks and WM declines with age. Event-related potentials (ERPs) evoked by certain tasks are also associated with specific cognitive functions. This study investigates whether specific types of working memory capacities are related to corresponding neural indices of attention and context updating (P3; visual oddball task) or semantic (N400; word-pair paradigm task) processes. In addition, it examines whether age affects this relationship. Younger adults (n=30; ages 18-23) and older adults …

Investigation Into The Potential For Acquired Cross Tolerances Of Amphetamines, Benjamin Weldon

CMC Senior Theses

In the United States, the treatment of ADHD through use of psychostimulants in adolescents is a growing reality that many people know today and many more will know in the coming years. Although the effectiveness of psychostimulants is a known asset, the benefits of these medications may not be a permanent solution, suggesting not only the presence of a tolerance build up, but as many stimulants are prescribed in a joint effort to combat ADHD, a possible cross tolerance between two prescribed medications. There are minimal studies published that have addressed the topic of cross stimulation tolerance buildup between prescription …

Extension Of The Ergot Alkaloid Gene Cluster, Samantha Joy Fabian

Graduate Theses, Dissertations, and Problem Reports (ETD)

Specialized metabolites produced by fungi impact human health. A large portion of the pharmaceuticals currently on the market are derived from metabolites biosynthesized by microbes. Ergot alkaloids are a class of fungal metabolites that are important in the interactions of environmental fungi with insects and mammals and also are used in the production of pharmaceuticals. In animals, ergot alkaloids can act as partial agonists or antagonists at receptors for 5-hydroxytryptamine (serotonin), dopamine, and noradrenaline as ergot alkaloids have chemical structures similar to those neurotransmitters. Therefore, they affect insects and mammals that consume them and can be used to produce drugs …

Identification And Functional Characterization Of Mads-Box Proteins And Ccos Involved In Carotenoid Metabolism In Durian Durio Zibethinus L. Pulps, Kittiya Tantisuwanichkul

Chulalongkorn University Theses and Dissertations (Chula ETD)

Durian (Durio zibethinus L.), known as the "King of fruits," holds significant economic importance in Thailand. Apart from its unique odor and sweet taste, durian pulp exhibits a yellow color, which is an important aspect of fruit quality. Carotenoids are essential compounds responsible for the yellow, orange, and red coloration in many fruits and vegetables. During durian fruit development and ripening processes, its pulp changes from white to yellow due to increased carotenoid accumulation. The carotenoid biosynthetic pathway, responsible for carotenoid production, is controlled by ethylene. However, the mechanism underlying this process in durian fruit is still unknow. Notably, the …

Producing Grnas For Use In Crispr-Cas9 Dna Editing Applications Via In Vivo T7 Rna Polymerase Transcription In Escherichia Coli., Christian A. Santiago

Honors Program Theses

The development of the CRISPR-Cas9 system is widely considered to be a breakthrough advancement in gene editing. Compared to traditional technologies, CRISPR is generally regarded as the most accessible, effective, and precise tool for inducing stable and inheritable gene edits in vivo. The CRISPR-Cas9 gene editing complex relies on the complementary binding of guide RNAs to specifically target a DNA sequence for endonuclease cleavage and mutagenesis. The production of gRNAs for use in CRISPR applications can become exceedingly complex and costly, as in vivo methodologies generally require the use of RNA polymerases that interfere with gRNA sequence identity and in …

Creating Meaningful Connections With The Electr Eating Meaningful Connections With The Electron Transport Chain Beyond A Virtual Classroom, Lesley Ann Giddings

Chemistry: Faculty Publications

At the center of feminist pedagogy is community. Creating community within the classroom and between the class and local communities increases student engagement and access to knowledge. Communities can be a source of solidarity and further decenter authority in the classroom, empowering students, teachers, and the surrounding community to be co-learners. Amid the COVID-19 pandemic, remote teaching posed challenges to student engagement. Here, I describe an assignment used to build an inclusive community of co-learners within a virtual biochemistry class and our local communities. Oxidative phosphorylation is a cellular process used to create energy that depends on the electron transport …

Detection Of Sars-Cov-2 Neutralizing Antibodies In Retropharyngeal Lymph Node Exudates Of White-Tailed Deer (Odocoileus Virginianus) From Nebraska, Usa, Korakrit Poonsuk, Duan S. Loy, Rachael Birn, Bryan Buss, Matthew Donahue, Todd Nordeen, Kylie Sinclair, Luke Meduna, Bruce Brodersen, John Dustin Loy

School of Veterinary and Biomedical Sciences: Faculty Publications

Disease surveillance testing for emerging zoonotic pathogens in wildlife is a key component in understanding the epidemiology of these agents and potential risk to human populations. Recent emergence of SARS-CoV-2 in humans, and subsequent detection of this virus in wildlife, highlights the need for developing new One Health surveillance strategies. We used lymph node exudate, a sample type that is routinely collected in hunter-harvested white-tailed deer (WTD, Odocoileus virginianus) for surveillance of chronic wasting disease, to assess anti-SARS-CoV-2 neutralizing antibodies. A total of 132 pairs of retropharyngeal lymph nodes collected from Nebraska WTD harvested in Nebraska, US, in 2019 …

Lead Induced Differences In Bone Properties In Osteocalcin +/+ And −/− Female Mice, G. Yildirim, W. C. Budell, O. Berezovska, S. Yagerman, S. S. Maliath, P. Mastrokostas, S. Tommasini, T. L. Dowd

Publications and Research

Lead (Pb) toxicity is a major health problem and bone is the major reservoir. Lead is detrimental to bone, affects bone remodelingand is associated with elderly fractures. Osteocalcin (OC) affects bone remodeling, improves fracture resistance and decreases with age and in some diseases. The effect of lead in osteocalcin depleted bone is unknown and of interest. We compared bone mineral properties of control and Pb exposed (from 2 to 6 months) femora from female adult C57BL6 OC+/+ and OC−/− mice using Fourier Transform Infrared Imaging (FTIRI), Micro-computed tomography (uCT), bone biomechanical measurements and serum turnover markers (P1NP, CTX). Lead significantly …

Characterizing Cellular Injury And Inflammation With Aldob-/-Mouse Model Of Mafld, Andy Rashid

All Master's Theses

Metabolic-associated fatty liver disease (MAFLD) affects 1 in 3 people in the US and is a healthcare burden on the order of billions of dollars. MAFLD is characterized by pathological lipid accumulation in the liver, excess production of triglycerides and cholesterol, and export of LDL particles that raise serum lipid levels. These metabolic perturbations can lead to the development of liver fibrosis, chronic inflammation, and insulin resistance. Diagnosing MAFLD prior to fibrosis is crucial for successful intervention and prevention of advanced liver disease and other metabolic complications. Fructose-1Phospahte (F1P) accumulation from fructose overconsumption can drive hepatic lipogenesis and MAFLD progression. …