Molecular Biology Commons^™

Development Of A Cure In Principles Of Cell And Molecular Biology, Heather Seitz

Sabbatical Projects

This project presentation highlights the work done during sabbatical to develop an authentic research experience into BIOL135 Principles of Cell and Molecular Biology. The sabbatical allowed for development time but further work was done to measure impact and outcomes for future implementation.

Effect Of Histidine Tag On The Enzymatic Activity Of Pseudolysin And Its Interaction With The Streptomyces Metalloprotease Inhibitor (Smpi) Protein, Adewale Adeboye Adewole

Electronic Theses and Dissertations

Pseudolysin (PLN) protein is a major virulent factor in Pseudomonas aeruginosa infection. PLN belongs to the family of zinc-dependent metalloproteases. It is naturally inhibited by a smaller protein called streptomyces metalloprotease inhibitor (SMPI), secreted by Streptomyces nigrescens. Previous and current studies demonstrate that the interaction between PLN and SMPI offers unique mechanistic insights for drug development against infections caused by P. aeruginosa. This study investigated the effect of N-terminal Histidine (His) tag on the enzymatic activity of PLN and its interaction with SMPI. His-tagged PLN and SMPI proteins were expressed in bacteria and purified by affinity chromatography …

Novel Nanomaterials For Developing Bone Scaffolds And Tissue Regeneration, Nazim Uddin Emon, Lu Zhang, Shelby Dawn Osborne, Mark Allen Lanoue, Yan Huang, Z. Ryan Tian

Cell and Molecular Biology Faculty Publications and Presentations

Nanotechnologies bring a rapid paradigm shift in hard and soft bone tissue regeneration (BTR) through unprecedented control over the nanoscale structures and chemistry of biocompatible materials to regenerate the intricate architecture and functional adaptability of bone. This review focuses on the transformative analyses and prospects of current and next-generation nanomaterials in designing bioactive bone scaffolds, emphasizing hierarchical architecture, mechanical resilience, and regenerative precision. Mainly, this review elucidated the innovative findings, new capabilities, unmet challenges, and possible future opportunities associated with biocompatible inorganic ceramics (e.g., phosphates, metallic oxides) and the United States Food and Drug Administration (USFDA) approved synthetic polymers, including …

Elucidating The Mechanisms Underlying A Functional Plant-Arbuscular Mycorrhizal Symbiosis Using Omics Approaches, Jasmine Gill

All Dissertations

Arbuscular mycorrhizal fungi (AMF) are obligate biotrophs forming symbiotic association with plants. The plant-AMF symbiosis has gained much attention due to its ability to improve plant nutrient uptake, especially phosphorus, which is often inadequately available to plants due to its limited mobility and complex formation in many soils. Phosphorus (P) is a crucial nutrient for plants, and most phosphate fertilizers are derived from phosphate rock, a finite natural resource. Thus, optimizing P uptake is vital for sustaining crop yields and addressing the needs of a growing global population. Plant-AMF symbioses are significant in this context, as AMF can acquire P …

Analysis Of An Ethanol Sensitive Bmp-Nkx2.3-Fgf Signaling Pathway In Pouch Morphogenesis., Hieu Dai Le Vo

Electronic Theses and Dissertations

Craniofacial malformations lie at the heart of Fetal Alcohol Spectrum Disorders (FASD). While there is growing evidence for a genetic component to FASD, little is known of the cellular mechanisms underlying these ethanol-sensitive loci in facial development. Bone Morphogenetic Protein (Bmp) signaling pathway dependent pouch formation is a key mechanism in facial development. We have previously shown that multiple Bmp mutants are sensitized to ethanol-induced facial defects. However, ethanol does not directly impact Bmp signaling. This suggests that downstream effectors, like nkx2.3 and Fibroblast Growth Factor (Fgf) signaling, may mediate the impact of ethanol on Bmp mutants. Here, I …

Exploring Variability Of The Cg18562578 Methylquant Assay For The Forensic Discrimination Of Monozygotic Twins, Sydney E. Arnold

Student Theses

Monozygotic (MZ) twins possess identical DNA sequences, posing a problem for forensic scientists, as they are incapable of distinguishing MZ twins from each other using traditional DNA identification methods. However, the epigenomes of MZ twins will differ and might be used for their forensic discrimination. Here, we explored the variability of an assay previously developed by our laboratory that showed high discrimination potential for MZ twins using buccal swab DNA, cg18562578 MethylQuant. We measured precision at each step: bisulfite conversion, nested PCR, and qPCR, reproducibility, and repeatability. DNA methylation ratios were compared between two analysts carrying each step using the …

Cg18562578 Methylquant Assay For The Discrimination Of Monozygotic Twins, Amber P. Chauca

Student Theses

Monozygotic (MZ) twins have identical DNA, making traditional profiling methods insufficient in their discrimination. However, MZ twins can be differentiated by their levels of DNA methylation. Our research group previously developed a methodology to detect cg18562578 DNA methylation levels in MZ twins based on a two- step PCR approach and a cg18562578 MethylQuant Assay. That study generated data for only 64% of the samples it investigated, and showed a discrimination power of 77% to discriminate between MZ twins. Here, we used the same methodology but a different approach to determine DNA methylation in 73 previously unsuccessful twin pairs. We diluted …

Biological, Histological, And Immunohistochemical Responses To Heat Stress In Common Goldfish, Carassius Auratus, Ngozichukwu Francis Ngozichukwu

Theses and Dissertations

Global climate change has led to rising temperatures, posing significant challenges to aquatic ecosystems and the health of ectothermic species such as goldfish (Carassius auratus) This thesis investigated the effects of heat stress on growth performance, histological alterations, and molecular stress responses in the gill and kidney tissues of goldfish Fish were exposed to control (24°C), medium (28°C), and high (32°C) temperatures for two weeks Growth parameters, including instantaneous growth rate (IGR), absolute growth rate (AGR), body value gain (BVG), and condition factor, were evaluated alongside histological and immunohistochemical markers of stress Results revealed that high temperature significantly reduced AGR …

Transcriptomic Study On The Effect Of Salt Stress In Sorghum Bicolor Seedlings, Joshwa A. Gandy

Theses and Dissertations

The global impact and reliance on cereal crops, and sorghum in particular, is paramount for worldwide food security and for the livelihood of hundreds of millions of people. The detrimental effects of salt stress on crop plants and the huge economic losses it causes by affecting agricultural output worldwide is a devastating issue with huge impacts, which have prompted research to combat salinity in numerous ways. Understanding how cereals respond to the negative effects of salt stress is crucial for enhancing their salt tolerance. In this study of sorghum, we used a 3’-end RNA sequencing approach to elucidate major salt …

Crystal Structures Of Escherichia Coli Glucokinase And Insights Into Phosphate Binding, Joseph Andrews, Joshua Sakon, Chenguang Fan

Chemistry & Biochemistry Faculty Publications and Presentations

Here, we report the crystal structure of Escherichia coli glucokinase (GLK), which has phosphate bound in the cleft between the alpha and beta domains adjacent to the active site. A ternary complex consisting of GLK, glucose and phosphate is also reported in this work. Diffraction data were collected at 2.63 angstrom resolution for the phospate-bound form (R-work/R-free = 0.191/0.230) and at 2.54 angstrom resolution for the ternary complex (R-work/R-free = 0.202/0.258), both at 297 K. A B-factor analysis of the phosphate-bound GLK structure revealed consistently lower values for phosphate-interacting basic residues in the alpha 4, alpha 5 and alpha 9 …

Metabolic Regulation Of Cardiac Fibroblast Phenotype And Function., Collin Kleis Wells

Electronic Theses and Dissertations

Cardiac fibroblasts are central effectors of cardiac repair after myocardial infarction (MI). In response to signaling cues, they differentiate to a range of phenotypes with robust capacities to synthesize and secrete extracellular matrix (ECM) and signaling molecules. Although activated fibroblast phenotypes are associated with pronounced changes in metabolism, it remains unclear how the metabolic network upholds the effector functions of fibroblasts in the post-infarcted heart. Here, we identified that critical enzymes in the phosphoenolpyruvate (PEP) cycle, i.e. pyruvate kinase muscle-2 (PKM2) and phosphoenolpyruvate carboxykinase-2 (PCK2), are elevated in the heart after MI and examined their role in regulating post-MI remodeling …

Use Of The Protein Misfolding Cyclic Amplification For Food Safety And Drug Discovery, Maria R. Benavente Garcia-Huidobro

Dissertations and Theses (Open Access)

Prion diseases are fatal neurodegenerative disorders caused by the misfolding of the normal prion protein (PrP^C) into its infectious form (PrP^Sc). While the zoonotic potential of chronic wasting disease (CWD) remains uncertain, the presence of prions in food products raises public health concerns. Additionally, therapeutic strategies for prion diseases remain limited. This thesis presents a methodological exploration of the Protein Misfolding Cyclic Amplification (PMCA) technique to address these challenges in two key areas: (1) detecting CWD prions in processed meats subjected to common cooking procedures, and (2) identifying potential anti-prion compounds through a high-throughput PMCA adaptation. …

Bifunctional Fusion Protein Pdl1sfv/Micae For Cancer Immunothearpy, Junyi Li

All Dissertations

Cancer immunotherapy has highlighted the importance of immune checkpoint blockade and innate immune cell engagement in battling tumor-mediated immune suppression in the past few decades. However, with cancer development, advanced tumors are often found to develop resistance towards single-target immunotherapy due to the complexity of the immunosuppressive mechanisms within the tumor microenvironment (TME). To address this, we developed a novel bifunctional fusion protein, PDL1sFv/MICAe, which combines the tumor-targeting capability of an anti-PDL1 single-chain variable fragment (sFv) with the NK cells immunostimulatory properties of the MICA extracellular domain.

In vitro functional assays revealed that PDL1sFv/MICAe significantly enhanced cytotoxicity towards natural killer …

Molecular And Clinical Insights Into Role Of Gper In Hepatocellular Carcinoma (Hcc), Hossain Ahmed

Theses and Dissertations

Metabolic dysfunction–associated steatotic liver disease (MASLD), encompassing stages from steatosis (MASLD) to steatohepatitis (MASH), cirrhosis, and hepatocellular carcinoma (HCC), is the most prevalent chronic liver disease worldwide, yet molecular markers of progression remain limited. The G protein–coupled estrogen receptor (GPER), a non-genomic estrogen receptor with metabolic and anti-inflammatory roles, emerges as a candidate regulator in hepatic pathophysiology. In this Dissertation thesis, we have delineated for the first time clinicopathological assessment of liver biopsies across MASLD stages revealing a robust inverse correlation between GPER expression and disease severity. Higher GPER levels in early MASLD associated with favorable metabolic and biochemical profiles, …

In Silico Prediction Of Cytotoxic T-Cell Epitopes From Helicobacter Pylori Virulence Factors Using An Immunoinformatics Approach, Demy Valerie Chacon, Kiana Alika Co, Daphne Noreen Enriquez, Aubrey Love Labarda, Reanne Eden Manongsong, Edward Kevin B. Bragais

Biology Faculty Publications

Background: Helicobacter pylori infects approximately half of the global population, leading to gastric and duodenal ulcers. Despite the availability of antibiotics, challenges such as patient reluctance, high treatment costs, and antibiotic resistance limit their effectiveness, making vaccination a promising alternative. This study used immunoinformatics to identify candidate epitopes for a multiepitope vaccine construct against H. pylori.

Material and methods: The protein variability server was utilized for conservation analysis. The epitopes were screened for antigenicity, allergenicity, toxicity, cross-reactivity, and population coverage. Selected epitopes were docked with their corresponding human leukocyte antigen (HLA) alleles, and thermodynamic quantities were determined. Five virulence …

The Influence Of Myocardin Effector Genes On The Tgf-Beta Smad3 Signaling Pathway In Human Pleural Mesothelial Cells, Hema Chandra Hrushi Dusharla

Biotechnology Theses

Pleural Fibrosis (PF) involves excessive extracellular matrix (ECM) deposition, mainly by myofibroblasts, leading to pleural thickening and impaired lung function. Pleural mesothelial cells (PMCs) contribute via mesothelial and mesenchymal transition (MesoMT), triggered by cytokines like TGF-β. Our lab identified Myocardin (MyoCD), a transcriptional co-activator of cardiac and smooth muscle, is a master regulator of pleural fibrosis through interactions with serum response factor (SRF) and Smad2/3 transcription factors which initiate fibrosis signaling. RNA sequencing and qPCR analysis revealed that silencing MyoCD decreased the expression of CTNNB1, MDFI, and Tspan2. MDFI emerged as a potent candidate, as this gene affecting the expression …

Using The Safety Scissors: Dna Resection Regulation At Dna Double-Strand Breaks And Telomeres, Michael M, Soniat, Logan R. Myler

Department of Biochemistry and Molecular Biology Faculty Papers

DNA resection is a universal process in genome maintenance by which one strand of DNA is degraded, leaving the other strand intact. This sometimes highly processive process is critical for many forms of DNA damage repair, replication-coupled repair, meiotic recombination, and telomere maintenance. Therefore, resection must be tightly regulated to prevent genome instability and promote faithful and accurate repair. Here, we review what is known about how resection functions and how it is controlled, using DNA double-strand break repair and telomere maintenance as examples. We address how resection is regulated in three independent steps: resection initiation, long-range processing, and termination. …

Elucidating The Role Of Mir-10a/B In The Wound Healing Process In Larval Zebrafish, Jacquelyn E. Jacobs

All NMU Master's Theses

This study explores the regulatory role of microRNA-10 (miR-10) in larval zebrafish caudal fin regeneration, a model with broad implications for the future of regenerative medicine. Building on prior research from the lab, which indicated that miR-10 knockdown impairs regeneration, we verified this finding through rescue experiments, assessed its influence on healing projections, and examined its role in the innate immune response during the wound healing process. To address these questions, we employed a multi-faceted approach, including timed matings, microinjections, immunohistochemistry, fixed sample microscopy, and live imaging. Our results show that the experimental introduction of exogenous miR-10 can rescue the …

From Differentiation To Disease: Cyclin C Directly Coordinates Transcription And Mitochondrial Dynamics, Alicia Campbell

Theses and Dissertations

Organelle crosstalk is essential for coordinating cellular responses to various external cues, including stress and differentiation. Two key organelles, the nucleus and mitochondria, engage in extensive crosstalk to manage responses to a range of environmental and developmental signals. Complex processes such as stem cell differentiation and neuronal development rely on organelle crosstalk. Both processes integrate changes in gene expression with cytosolic and organellar remodeling. However, it remains unclear how these two complex processes are coordinated to achieve the desired outcome. This project concentrates on cyclin C (CCNC), a component of the Mediator Kinase Module (MKM), and its role in directly …

The Structure Of The Drp1 Lattice On Membrane, Kristy Rochon, Anelise N. Hutson, Jason A. Mears

Faculty Scholarship

Mitochondrial health relies on the membrane fission mediated by dynamin-related protein 1 (Drp1). Previous structural studies of Drp1 on remodeled membranes were hampered by heterogeneity, leaving a critical gap in the understanding of the mitochondrial fission mechanisms. Here we present a cryo-electron microscopy structure of full-length human Drp1 decorated on membrane tubules. Using the reconstruction of average subtracted tubular regions (RASTR) technique, we report that Drp1 forms a locally ordered lattice along the tubule without global helical symmetry. The filaments in the lattice are similar to dynamin rungs with conserved stalk interactions. Adjacent filaments are connected by GTPase domain interactions …