Biochemistry, Biophysics, and Structural Biology Commons^™

Structural And Functional Studies Of Ascc1 In Dna And Rna Alkylation Damage Response, Naga Babu Chinnam

Dissertations and Theses (Open Access)

Alkylating agents from environmental, endogenous, and certain chemotherapeutic sources constantly challenge cellular nucleic acids. Repairing the damage caused by alkylation is necessary for maintaining genomic stability. ALKB family Human Homolog ALKBH3 plays a key in repairing alkylated damages in DNA and RNA molecules. Activating signal co-integrator complex 1 (ASCC1) acts with ASCC-ALKBH3 complex in alkylation damage responses. ASCC1 uniquely combines two evolutionarily ancient domains: nucleotide-binding K-Homology (KH) (associated with splicing, transcriptional regulation, and translational control) and two-histidine phosphodiesterase (PDE) (associated with hydrolysis of cyclic nucleotide phosphate bonds). Germline mutations link ASCC1 loss of function to spinal muscular atrophy with congenital …

Testing Resilience Of Microalgae Biofilms When Exposed To Hydrothermal Liquefaction Aqueous Phase, Davis Riley Haag

Undergraduate Honors Capstone Projects

There are a multitude of experimental and processing techniques in the bioenergy field that produce sustainable and carbon negative to carbon neutral methods of converting renewable resources into biologically based hydrocarbons. One example of a process is hydrothermal liquefaction (HTL) which specializes in converting wet biomass into biocrude oil. One reason that these processes are not used outside experimental testing is due to the high cost of performing these processes. There is potential to move these processes to larger scale testing and eventually industrial adaptation if breakthroughs are made to offset the process expenses. There is a waste product produced …

Exploring Novel Nutrient Acquisition Mechanisms For Maintaining Energy Metabolism In Toxoplasma Gondii, Melanie Key

All Dissertations

Toxoplasma gondii is an infectious intracellular parasite that establishes infection within various mammalian hosts, including humans. Toxoplasma infects approximately one-third of the global population, making it an impactful pathogen of study. T. gondii is the causative agent of toxoplasmosis, an opportunistic infection that causes severe illness or fatality in immunocompromised individuals or pregnant women. The parasite possesses a very effective invasion strategy, allowing it to cross different tissue types to disseminate infection across the placental and blood-brain barrier. Although there are a few treatments for acute toxoplasmosis, these drugs can produce adverse side effects. These options are not effective against …

Exploring The Effects Of N,N-Dimethylbiguanide And Ω-3 Pufas On Muscle Stem Cell Differentiation, Enayah Alhshim

Graduate Theses and Dissertations

The impacts of omega-3 (ω-3) fatty acid and metformin hydrochloride (Met-HCl) on muscle stem cells differentiation, mimicking the fetal muscle development under gestational diabetes mellitus (GDM), were investigated. ω-3 fatty acids have been known for their ability to regulate adipogenic transcription factors such as Sterol Regulatory Element-Binding Protien (SREBPs) and Peroxisome Proliferator-Activated Receptors (PPARs). This regulation resulted in gene expression modification for lipid metabolism and adipocyte differentiation, thus preventing excessive fat accumulation. Additionally, emerging evidence suggested that ω-3 fatty acids may also exert direct effects on myogenic processes that influence muscle stem cells proliferation, differentiation, and maturation. Meanwhile, Met-HCl, an …

Comparing The Regulatory Effects Of Overexpressed Micrornas And Xenobiotic Drugs On Cell Cycle And Apoptotic Regulators In Pc-3 Cells, Tommie Johnson

Dissertations (2016-Present)

MicroRNA was first discovered in C. elegans as small temporal RNA (stRNA) that does not code for protein. Since being discovered they have played a significant role in regulating gene expression at the post-transcriptional level. miRNAs are found in various organisms, and they bind to the 3' untranslated regions to inhibit translation and cause mRNA degradation. Some drugs are involved in cell cycle regulation such as Palbociclib, Ribociclib (LEE011), and Abemaciclib (LY2835219), that causes G1 arrest impeding cell proliferation. Cyclin D1 (CCND1) is supported by the cell cycle making it into a functional product. When undergoing a chemical reaction, the …

The Expression Of Recombinant Cas9 Proteins In E. Coli Bl21(De3) Strains, Morgan Reese

Graduate Theses and Dissertations

Abstract Extensively used for genome editing, CRISPR-Cas9 is a relatively new tool that has made a major impact in the genomic engineering landscape. Due to this, the production of CRISPR proteins that are not commercially available can limit research applications. Here, we present methods for optimizing laboratory scale production of CRISPR proteins including Cas9. The method uses IPTG to induce gene expression of the specified Cas9 and nickel column purification to isolate proteins. CRISPR proteins are validated for purity by SDS-PAGE gel, western blot, and for activity by in vitro digestion. We thoroughly validate and compare the effectiveness of BL21(DE3) …

Simvastatin Induces Autophaghy-Mediated Cell Death In Metastatic Breast Cancer Cells, Jessica Allagoa

Theses (2016-Present)

Breast cancer remains one of the leading causes of cancer deaths among women. Due to the limited effectiveness of current anticancer drugs, ongoing research has extended towards alternative drug categories for potential treatments. Recent findings indicate that statins possess the ability to suppress tumors across various cell types. Traditionally, statins are known as a class of cholesterol-lowering agents and function by inhibiting 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase, a key enzyme in the mevalonate pathway. However, statins can also suppress cell proliferation and ultimately lead to cell death, which includes Type I apoptosis-induced cell death or Type II autophagy-induced cell death. Autophagy is …

Foxo1 Inhibitor, As1842856, Induces Cell Cycle Arrest And Reverses Anticancer Drug-Induced Cytotoxicity In Osteosarcoma Cells, Antanay Hall

Theses (2016-Present)

Forkhead box class O (FOXO)-1 transcription factor controls cell proliferation, apoptosis, oxidative stress, and other cellular activities; FOXO1 has also been implicated in cell cycle regulation. This research project aims to determine the contribution of FOXO1 to cell cycle regulation and response to anticancer drug treatment in osteosarcoma. Osteosarcoma is the most common bone cancer, with most cases occurring in people younger than 30 years old. The study explores the impact of FOXO1 inhibitor AS1842856 on the cytotoxic effects of anticancer drugs in CCHOSD, Hos, and LM7 osteosarcoma (OGS) cell lines. Following chemical inhibition of FOXO1 and anticancer drug treatment, …

A Combination Of Generated Hydrogen Sulfide And Nitric Oxide Activity Has A Potentiated Protectant Effect Against Cisplatin Induced Nephrotoxicity, Faria Khurshid, Javeid Iqbal, Fiaz Ud Din Ahmad, Arslan Hussain Lodhi, Abdul Malik, Suhail Akhtar, Azmat Ali Khan, Marvi Imam Bux, Mohammed Younis

Biomedical Sciences Faculty Publications

Aim: Hydrogen sulfide and nitricoxide possess cytoprotective activity and in vivo, they are generated from exogenous sodium hydrosulfide and L-arginine respectively. Cisplatin is a major chemotherapeutic agent used to treat cancer and has a high incidence of nephrotoxicity as a side effect. The study aim was to explore the effects of NaHS and L-arginine or their combination on cisplatin induced nephrotoxicity in rats. Methods: Wistar Kyoto rats were given a single intraperitoneal dose of cisplatin (5 mg/kg) followed either by NaHS (56 μmol/kg, i. p.), L-arginine (1.25 g/L in drinking water) or their combination daily for 28-days. Post-mortem plasma, urine …

Molecular Insights Of Paraoxonase 2 In Advanced Glycation End Product Induced Endothelial Dysfunction, Ramya R

Theses and Dissertations

Diabetes mellitus (DM) is a metabolic disorder associated with various vascular complications and affects the function of the endothelial cells. Elevated blood glucose levels accelerate the formation of advanced glycation end products (AGEs) by Amadori rearrangement and bind to their Receptor for the advanced glycation end product (RAGE) and evoke release of key pro-inflammatory cytokine proteins leading to endothelial dysfunction. Paraoxonase 2 (PON2) is an endogenous intracellular enzyme, localised in mitochondria, endoplasmic reticulum (ER), and nuclear membrane with numerous properties, like anti-inflammatory, anti-apoptotic and anti-oxidant.

Our present study aims to ascertain the effect of AGEs on PON2 activity and expression, …

Overexpression Of Fatty Acid Synthase Upregulates Glutamine–Fructose-6-Phosphate Transaminase 1 And O-Linked N-Acetylglucosamine Transferase To Increase O-Glcnac Protein Glycosylation And Promote Colorectal Cancer Growth, James Drury, Mariah E. Geisen, Josiane Weber Tessmann, Piotr G. Rychahou, Courtney O. Kelson

Markey Cancer Center Faculty Publications

Fatty acid synthesis has been extensively investigated as a therapeutic target in cancers, including colorectal cancer (CRC). Fatty acid synthase (FASN), a key enzyme of de novo lipid synthesis, is significantly upregulated in CRC, and therapeutic approaches of targeting this enzyme are currently being tested in multiple clinical trials. However, the mechanisms behind the pro- oncogenic action of FASN are still not completely understood. Here, for the first time, we show that overexpression of FASN increases the expression of glutamine–fructose-6-phosphate transaminase 1 (GFPT1) and O-linked N-acetylglucosamine transferase (OGT), enzymes involved in hexosamine metabolism, and the level of O-GlcNAcylation in vitro …

The Purification And Thermal Stability Of The Peroxidase Enzyme In Cucurbita Moschata, Garen Hamner

Senior Honors Theses

Peroxidases are enzymes that catalyze the reduction of hydrogen peroxide to water while oxidizing organic substrates and are valuable in spheres like industrial and medical applications and histochemistry. Limitations still exist in the use of the well-studied horseradish peroxidase for certain activities due to limitations like poor thermal stability, thus the search for novel peroxidases that can overcome these limitations is an active area of research. Butternut squash peroxidase (Cucurbita moschata) (BSP) shows promise due to significant activity being found in the skin and apparent enhanced thermal stability, but an efficient purification scheme for it is lacking, as well as …

Using Edna To Assess Impacts Of Oyster Restoration On Ecosystem Biodiversity At A Heavily Impacted Coastal Lagoon, Dominique Di Domenico, Emily Bonacchi, Elizabeth Suter

Molloy Multidisciplinary Undergraduate Research Conference

The Great South Bay (GSB), a bar-built, temperate, coastal lagoon on the south shore of Long Island, New York, USA, is a highly modified estuary due to heavy urbanization and suburbanization in the last century. GSB historically provided up to 50% of the nation’s hard clam (Mercenaria mercenaria) harvest, and is the legal home of the “Blue Point” oyster (eastern oyster; Crassostrea virginica). However, due to overexploitation as well as persistent stress from ongoing nitrogen pollution and harmful algal blooms, shellfish populations are fractions of their original abundances. Restoration efforts in GSB, driven both by community efforts …

Trna Anticodon Cleavage By Target-Activated Crispr-Cas13a Effector, Ishita Jain, Matvey Kolesnik, Konstantin Kuznedelov, Leonid Minakhin, Natalia Morozova, Anna Shiriaeva, Alexandr Kirillov, Sofia Medvedeva, Alexei Livenskyi, Laura Kazieva, Kira S Makarova, Eugene V Koonin, Sergei Borukhov, Konstantin Severinov, Ekaterina Semenova

Rowan-Virtua School of Osteopathic Medicine Departmental Research

Type VI CRISPR-Cas systems are among the few CRISPR varieties that target exclusively RNA. The CRISPR RNA–guided, sequence-specific binding of target RNAs, such as phage transcripts, activates the type VI effector, Cas13. Once activated, Cas13 causes collateral RNA cleavage, which induces bacterial cell dormancy, thus protecting the host population from the phage spread. We show here that the principal form of collateral RNA degradation elicited by Leptotrichia shahii Cas13a expressed in Escherichia coli cells is the cleavage of anticodons in a subset of transfer RNAs (tRNAs) with uridine-rich anticodons. This tRNA cleavage is accompanied by inhibition of protein synthesis, thus …

Structural Homology Of Mite Profilins To Plant Profilins Is Not Indicative Of Allergic Cross-Reactivity, Andrea O'Malley, Sahana Sankaran, Avery Carriuolo, Kriti Khatri, Krzysztof Kowal, Maksymilian Chruszcz Ph.D.

Faculty Publications

Structural and allergenic characterization of mite profilins has not been previously pursued to a similar extent as plant profilins. Here, we describe structures of profilins originating from Tyrophagus putrescentiae (registered allergen Tyr p 36.0101) and Dermatophagoides pteronyssinus (here termed Der p profilin), which are the first structures of profilins from Arachnida. Additionally, the thermal stabilities of mite and plant profilins are compared, suggesting that the high number of cysteine residues in mite profilins may play a role in their increased stability. We also examine the cross-reactivity of plant and mite profilins as well as investigate the relevance of these profilins …

Investigating The Role Of Tryptophan In The Native Photo-Crosslinking Of A Cys-Containing Transmembrane Helix To A Membrane Lipid Double Bond, Michael Jay Telehany

Undergraduate Honors Theses

Our research group has previously discovered that a Cys residue within a transmembrane (TM) helix can directly photo-crosslink to a membrane lipid via an oxythiolation addition reaction to the double bond within the lipid, with a reaction yield of 10-20%. The pH-low Insertion Peptide (pHLIP) serves as our model TM helix, while the POPC lipid bilayer is our model membrane. We had long believed that the Trp residues within the sequence of pHLIP served as the key chromophore for this reaction, and thus they were essential for the formation of the TM helix-lipid adduct. To test this hypothesis, I synthesized …

Modeling And Synthesis Of 3-(2,6-Disubstituted 5-Pyrimidyl) Propionic Acids As Inhibitors Of Low Molecular Weight Protein Tyrosine Phosphatase (Lmw-Ptp), Kennedy Bueno, Madisen Carter

Celebrating Scholarship and Creativity Day (2018-)

Low molecular weight protein tyrosine phosphatase (LMW-PTP) is an enzyme and a known signal pathway for growth factors and cellular transformation in eukaryotic cells. Our research aims to synthesize 12 new potential inhibitors of LMW-PTP and analyze their inhibitory activity and binding affinity. The inhibition of both isoforms of LMW-PTP has been the primary focus of research due to their potential role in breast, colon, and other cancers as well as type II diabetes. One known inhibitor, pyridoxal 5’- phosphate (PLP), is essential for various enzymatic reactions within the body, such as the synthesis of neurotransmitters, and is therefore impractical …

Mycobacteriophage Lberry, An A3 Cluster Phage Infecting Mycobacterium Smegmatis, Nathan Berry '25

Chemistry & Biochemistry Student Scholarship

Nathan Berry ’25, Biochemistry major
Faculty Mentor: Dr. Kathleen Cornely, Chemistry and Biochemistry

Synthesis Of 2-Substituted Beta-Carbolines From N-Oxides, Alma Martinez '26

Chemistry & Biochemistry Student Scholarship

Alma Martinez ’26, Biochemistry major
Faculty Mentor: Dr. Seann Mulcahy, Chemistry and Biochemistry

Mycobacteriophage Maravista, An F1 Cluster Phage Infecting Mycobacterium Smegmatis, Charles Pelagalli '25

Chemistry & Biochemistry Student Scholarship

Charles Pelagalli ’25, Biochemistry major
Faculty Mentor: Dr. Kathleen Cornely, Chemistry and Biochemistry

Mycobacteriophage Rita, Anna Fakhri '24

Chemistry & Biochemistry Student Scholarship

Anna Fakhri ’24, Biochemistry major
Faculty Mentor: Dr. Kathleen Cornely, Chemistry and Biochemistry

Mycobacteriophage Pembroke, An A3 Cluster Phage Infecting Mycobacterium Smegmatis, Marly Cassford '25

Chemistry & Biochemistry Student Scholarship

Marly Cassford ’25, Biochemistry major
Faculty Mentor: Dr. Kathleen Cornely, Chemistry and Biochemistry

Improving The Drug Antipyrine: Synthesis Of N2-Aryl Analogs Through Oxidation Of Dihydroantipyrines, Eli Mans

Student Academic Conference

Idiopathic Pulmonary Fibrosis (IPF) is a progressive and fatal lung disease that affects thousands of people worldwide. Current drugs are used in clinical settings to treat IPF but these drugs have low efficacy and a high monetary costs. The molecule antipyrine has been identified by colleagues at Mayo Clinic as an early-stage drug candidate for treatment for IPF. The Jasperse group is synthesizing analogs of antipyrine to create a drug library for improved IPF treatment. A novel three step process via a silyl enol ether intermediate has been developed to convert a C-C bond to a C=C bond between C4-C5 …

Predictive And Prognostic Biomarkers And Tumor Antigens For Targeted Therapy In Urothelial Carcinoma, Aditya Eturi, Amman Bhasin, Kevin Zarrabi, William Tester

Department of Medical Oncology Faculty Papers

Urothelial carcinoma (UC) is the fourth most prevalent cancer amongst males worldwide. While patients with non-muscle-invasive disease have a favorable prognosis, 25% of UC patients present with locally advanced disease which is associated with a 10-15% 5-year survival rate and poor overall prognosis. Muscle-invasive bladder cancer (MIBC) is associated with about 50% 5 year survival when treated by radical cystectomy or trimodality therapy; stage IV disease is associated with 10-15% 5 year survival. Current therapeutic modalities for MIBC include neoadjuvant chemotherapy, surgery and/or chemoradiation, although patients with relapsed or refractory disease have a poor prognosis. However, the rapid success of …

Synthesis Of Novel Temozolomide Amides As Potential Anti-Glioblastoma Agents, Sion Kim

Honors Theses

Glioblastoma is treated with Temozolomide (TMZ), a lipophilic alkylating drug that can cross the brain-blood barrier. To confront drug resistance, this research aimed to synthesize novel TMZ amide hybrids by using the acid chloride method, replacing an amino group with a primary amine. These novel hybrids were tested on U87-MG cells to determine if the primary amines with greater anticancer properties and lipophilicity have enhanced the drug's anticancer activity and bioavailability. The calculated lethal concentration 50 (LC50) of the hybrids ranged from 0.005 mg/mL to 0.2258 mg/mL, with some demonstrating greater anticancer effects than TMZ itself.

Isolation And Analysis Of R130d Malate Dehydrogenase (Mdh), Hayley Brunette, Abby Doyle, Ilsa Hoaglund, Danielle Bolland

Undergraduate Research Symposium 2024

Malate Dehydrogenase (MDH) is the enzyme that catalyzes the ninth step of the citric acid cycle, a regenerative step in the metabolism of glucose in which malate is converted to oxaloacetate. Conformational changes in the active loop region of MDH along with the coenzyme NAD+ drive catalysis. Previous studies suggested that mutating positively charged amino acids to negatively charged amino acids at residue 130 decreased the enzymatic activity of MDH. We hypothesized that if we change arginine 130 (positively charged amino acid) to aspartate (negatively charged amino acid), then the enzymatic activity of MDH would significantly decrease. In e. Coli …

Discovery Of Strong 3-Nitro-2-Phenyl-2h-Chromene Analogues As Antitrypanosomal Agents And Inhibitors Of Trypanosoma Cruzi Glucokinase, Shane M. Carey, Destiny M. O’Neill, Garrett B. Conner, Julian Sherman, Ana Rodriguez, Edward L. D'Antonio

Natural Sciences Faculty Publications

Chagas disease is one of the world’s neglected tropical diseases, caused by the human pathogenic protozoan parasite Trypanosoma cruzi. There is currently a lack of effective and tolerable clinically available therapeutics to treat this life-threatening illness and the discovery of modern alternative options is an urgent matter. T. cruzi glucokinase (TcGlcK) is a potential drug target because its product, D-glucose-6-phosphate, serves as a key metabolite in the pentose phosphate pathway, glycolysis, and gluconeogenesis. In 2019, we identified a novel cluster of TcGlcK inhibitors that also exhibited anti-T. cruzi efficacy called the 3-nitro-2-phenyl-2H-chromene analogues. This was achieved by performing a target-based …

Upregulation Of The Predominant Cystic Fibrosis Causing Mutation Df508-Cftr By Triazole Compounds In Epithelial Cells, Maggie Taylor

Undergraduate Research Conference

Cystic fibrosis is a common genetic disease that is caused by a mutation in the plasma membrane protein CFTR, which stands for Cystic Fibrosis Transmembrane-conductance Regulator. When this membrane protein is mutated, it impairs its chloride ion channel function, blocking the movement of chloride ions that travel in and out of the cell. Previous studies have demonstrated that the most prevalent CFTR mutation, ∆F508-CFTR, can be partially reversed using small molecules (Heda and Marino, BBRC, 271:659-664, 2000). In this study, I have investigated the effects of several triazole compounds known to bind and transport chloride ions in cultured cells, on …

Hgs-3 The Influence Of A Tandem Cycling Program In The Community On Physical And Functional Health, Therapeutic Bonds, And Quality Of Life For Individuals And Care Partners Coping With Parkinson’S Disease, Leila Djerdjour, Jennifer L. Trilk

SC Upstate Research Symposium

Purpose Statement: Several studies have shown that aerobic exercise can have a positive impact on alleviating symptoms experienced by individuals with Parkinson's disease (PD). Despite this evidence, the potential benefits of exercise for both PD patients and their care partners (PD dyad) remain unexplored. This research project investigates the effectiveness, therapeutic collaborations, and physical outcomes of a virtual reality (VR) tandem cycling program specifically designed for PD dyads.

Methods: Following approval from the Prisma Health Institutional Review Board, individuals with PD were identified and screened by clinical neurologists. The pre-testing measures for PD dyads (N=9) included emotional and cognitive status …

Lysine 473 Regulates The Activity And Trafficking Of The Cystine/Glutamate Transporter, System XC-, Anna Koppin

23rd Annual A. Paul and Carol C. Schaap Celebration of Undergraduate Research and Creative Activity (2024)

System x_c^- is a membrane transport system that plays a critical role in mitigating oxidative stress. Past work in our lab has shown that System x_c^- localizes to the plasma membrane allowing for increased activity to support production of antioxidants during oxidative stress. In this study, we sought to determine if post-translational modification (PTM) of the transporter regulates its trafficking. A C-terminal 3KR mutant (K422,472,473R) exhibited decreased membrane localization and activity, suggesting that PTM at one of these sites increases activity. Further, we observed that K473R exhibits a 7 kD decrease in the molecular weight, indicating …