Biotechnology Commons^™

Uti, Delirium, And Dementia: A Conceptual Bioinformatics Framework For Clinical Attribution, Rob E. Carpenter, Andrew Krouse

Human Resource Development Faculty Publications and Presentations

Urinary tract infection (UTI) is frequently invoked to explain delirium, confusion, and functional decline in persons with dementia. Yet urine positivity in this population often reflects asymptomatic bacteriuria, colonization, chronic pyuria, contamination, or sampling noise rather than clinically attributable infection. For biomedical informatics and computational diagnostics readers, this narrative review contributes an integrative conceptual bioinformatics framework for probabilistic clinical attribution of suspected UTI in dementia under uncertainty. We formalizes the attribution problem by distinguishing organism detection from a clinically plausible infection-related contribution to neurocognitive decline. The framework integrates clinical, laboratory, biological, and longitudinal data to clarify whether urinary findings are …

Virus-Like Particle Peptoid Conjugate As A Triple-Negative Breast Cancer Targeting Delivery System, Ricky Zhu

Chemistry Theses

Triple-negative breast cancer (TNBC), an aggressive form of breast cancer, characterized by its lack of estrogen receptors (ER), progesterone receptors (PR), and human epidermal growth factor receptors (HER2) poses a therapeutic challenge as common treatment options are rendered ineffective. Here is a strategy to develop a drug delivery system utilizing poly-glycinated LC129 8, a TNBC targeting peptoid, covalently attached to HK97 virus-like particles (VLP) containing a C-terminal LPETG sequence via a sortase mediated reaction. Using the properties of the cage-like structure of VLPs and the TNBC specificity of the LC129-8, high doses of chemotherapy can be efficiently loaded into the …

Thrombin-Induced Myofibroblast Differentiation Is Regulated By Myocardin And Novel Effector Genes In Ipf Fibroblasts, Christiana Okeke

Biotechnology Theses

Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease characterized by fibroblast activation, excessive extracellular matrix (ECM) deposition, and irreversible respiratory decline. In this study, we investigated the role of myocardin and its downstream effectors, RGS5, DLGAP5, and TAGLN, in thrombin-induced myofibroblast differentiation and ECM remodeling using primary human IPF fibroblasts. Myocardin knockdown significantly attenuated thrombin-induced expression of profibrotic genes, including FN1, COL1A1, PAI-1, ACTA2, and CNN1, as well as the novel effectors RGS5, DLGAP5, and TAGLN, confirming myocardin as a central regulator of fibrotic signaling. Silencing RGS5 produced minimal effects on gene and protein expression despite minor downward …

Myocardin-Dependent Regulation Of Adam12 And Adam19 Promotes Fibronectin Fibrillogenesis During Pleural Fibrosis, Uday Siva Naga Sankar Allamneni

Biotechnology Theses

Pleural fibrosis involves excessive extracellular matrix (ECM) deposition driven by chronic injury and inflammation. Transforming growth factor-β (TGF-β) induces mesothelial–mesenchymal transition (MesoMT), promoting a myofibroblast phenotype. Our laboratory identified myocardin (Myocd), a transcriptional coactivator, as a key mediator of MesoMT that regulates ECM-remodeling proteases. RNA-seq and qPCR analyses showed that ADAM12 and ADAM19 are upregulated by TGF-β but suppressed by Myocd knockdown in Human pleural mesothelial and IPF-derived cells. Silencing ADAM12 reduced fibrotic markers, including fibronectin and smooth muscle actin (α-SMA), whereas ADAM19 showed relatively minor effects. ADAM12 knockdown also impaired fibronectin fibrillogenesis and insoluble matrix assembly, as confirmed by …

Role Of Kif4a In The Fxa-Induced Phenotypic Modulation Of Pleural Mesothelial Cells, Sudishna Khanal

Biotechnology Theses

Pleural injuries can disrupt the organization of the pleural surface, leading to pleural remodeling and fibrosis (PF), characterized by the proliferation of myofibroblasts expressing alpha-smooth muscle actin (α-SMA) and the deposition of extracellular matrix proteins, primarily due to the mesenchymal transition of pleural mesothelial cells (PMCs) (MesoMT). It is well established that various external stimuli, such as TGF-β, thrombin, and factor Xa (FXa), induce MesoMT. Our laboratory found that myocardin, a transcriptional co-activator, was dramatically up-regulated during MesoMT, and that gene silencing of myocardin (MyoCD) diminished MesoMT. RNA sequencing data showed that FXa activated a set of genes not upregulated …

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 …

Effects Of Fibrogenic And Non Fibrogenic Nanomaterials On Cellular And Molecular Responses In Thp-1 Macrophage, Adedayo E. Oyerinde

Biotechnology Theses

Traditional toxicological testing of nanomaterials is often low-throughput and time-consuming. Here, we aimed to develop an economically viable in vitro assay to prescreen nanomaterials with fibrogenic potential. To this end, we tested the generation of reactive oxygen species (ROS) as well as cytokines (pro- and anti-inflammatory) by macrophages following treatment with fibrogenic and non-fibrogenic nanomaterials. Both classes of nanomaterials induced ROS indistinguishably, suggesting that ROS production doesn’t predict nanomaterial fibrogenicity. We observed higher levels of IL1β and IL6 in macrophages treated with fibrogenic nanomaterials. Analysis of single-cell RNA-Sequencing data of fibrotic lungs from rodents and humans revealed overexpression of Brain …

Engineering Hiv-1 Env Protein To Elucidate In Situ Structural Dynamics Of Env, Narendra Kumar Gonepudi

Biotechnology Theses

The HIV-1 Envelope (Env) glycoprotein trimers undergo structural changes upon interacting with host receptor CD4 and coreceptors (CCR5/CXCR4) for virus entry. As the only surface-exposed viral protein, Env is a key antibody target but evades recognition through structural changes. Studying Env dynamics with respect to the coreceptor binding region (V3 loop) provides insights into virus entry and immune evasion. Fluorescence labeling via genetic code expansion (amber–TAG suppression) offers a minimally invasive approach for fluorescence microscope-based Env dynamics studies. Here, two V3-related dual-amber Env constructs (A135_TAG-P308_TAG, P308_TAG-E395_TAG) were generated by incorporating two non-canonical …

Myocardin Effector Proteins Affect Tgf-Beta-Induced Myofibroblast Differentiation In Ipf Cells, Allison Gonzalez

Biotechnology Theses

Idiopathic pulmonary fibrosis (IPF) is an incurable interstitial lung disease defined by the progressive, irreversible scarring of the tissues between the lung’s alveoli. To date, only two drugs have received approval for treatment. However, these medications are not considered cures since they do not stop the progression of the disease. A deeper understanding of the mechanisms underlying IPF could aid in discovering a cure. We recently found that myocardin (MyoCD) regulates SMAD2/3 transcription factors in the TGF-β signaling pathway and governs the genes involved in pleural fibrosis. However, it is unclear whether MyoCD plays a critical role in IPF by …

Characterization Of The Slc7a5/Slc3a2 Amino Acid Transporter In Human Pleural Mesothelial Cells, Ann Francis

Biotechnology Theses

The SLC7A5/SLC3A2 amino acid transporter moves essential amino acids, such as leucine, phenylalanine, and tryptophan, across the cellular plasma membrane. SLC7A5 overexpression was identified in diverse cancers, suggesting a role in tumorigenesis and cancer cell proliferation. However, the role of SLC7A5/SLC3A2 in other aggressive cellular phenotypes, including mesothelial mesenchymal transition (MesoMT), remains unexplored. RNA sequencing analyses identified SLC7A5 as one of three highly upregulated SLCs in mesothelial cell derived myofibroblasts. This sodium independent transporter exhibits a high affinity for branched chain amino acids such as leucine; however, what role, if any, this channel plays in MesoMT is unclear. SLC7A5 knockdown …

Novel Role And Mechanisms Of Brd4 In Mediating Mesothelial To Mesenchymal Transition And Pleural Fibrosis, Felix Kankam

Biotechnology Theses

Pleural fibrosis is characterized by myofibroblast accumulation and increased expression of collagen and other extracellular matrices due to injury to the pleura and remodeling. Mesothelial to mesenchymal transition (MesoMT) is an important pathological event that contributes to the pool of myofibroblasts although its regulation remains incompletely understood. Recent research has reported the role of BRD4, a member of the bromodomain extra terminal domain (BET) family, in developing fibrosis in the heart and the lungs. One recent study showed that MEOX1 is a downstream target of BRD4 and mediated cardiac fibroblast activation. Our joint work also demonstrated that NOX4 is implicated …

Effect Of Monoclonal Antibodies Ma-33h1f7 And Ma-8h9d4 On Pai-1 In Cells Activated With Tgfβ, Srikar Modukuri

Biotechnology Theses

This study investigates the impact of monoclonal antibody treatment on plasminogen activator inhibitor-1 (PAI-1) expression and secretion in transforming growth factor beta (TGFβ) activated pleural human mesothelial cells. In this study, we examined total PAI-1 levels in conditioned media from human pleural mesothelial cells under various conditions using immunoblot and enzyme-linked immunosorbent assay techniques (ELISA). Specifically, we targeted PAI-1 with monoclonal antibodies (mAbs) MA-33H1F7 and MA-8H9D4, which bind to unique epitopes on PAI-1. A reduction in PAI-1 levels was observed between control and antibody-treated cells by immunoblot analysis of cell conditioned media. ELISA results also revealed a reduction in total …

Effects Of Interleukin-22 (Il-22) On Necroptosis, Inflammatory Responses, And Metabolism Of Mycobacterium Tuberculosis Stimulated Type 2 Diabetes Mellitus Mouse Macrophages, Bismark Owusu-Afriyie

Biotechnology Theses

Mycobacterium tuberculosis (Mtb) infects one-third of the world’s population and causes nearly 1.3 million deaths per year. Limited information is available about the immune responses during Mtb infection in type 2 diabetic hosts. Our laboratory developed an experimentally induced type 2 diabetes (T2DM) model in wild-type C57BL/6 mice and found that IL-22 and type 3 innate lymphoid cells (ILC3s) reduce inflammation and mortality of Mtb-infected T2DM mice. Our laboratory also found that Mtb-infected alveolar macrophages (AMs) from T2DM mice undergo necroptosis compared to Mtb-infected AMs of non-T2DM mice. In the current study, we determined whether …

The Role Of Myocardin In The Progression Of Non-Small Cell Lung Cancer, Soromidayo Akinsiku

Biotechnology Theses

Lung cancer is the leading cause of cancer-related mortality in the world and NSCLC accounts for 85% of all lung cancer cases. The mainstay of treatment for patients with stage I, II and IIIA NSCLC is surgery, followed by post-operative cisplatin-based chemotherapy. Additional adjuvant therapy involving targeted tyrosine kinase inhibitors has been in use, however even for the targeted therapy, resistance eventually develops. Therefore, there is a need for identifying novel targets for this life-threatening disease. Given that preliminary studies in Ikebe lab revealed that myocardin knockdown significantly promoted caspase-3 degradation, in this study, using myocardin siRNA, we investigated the …

Role Of Histone Deacetylases In Regulation Of Macrophage Phagocytosis Of Mycobacterium Tuberculosis, Benjamin Esiesimua

Biotechnology Theses

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a global health emergency. Macrophages are critical in protection against TB by phagocytosis of Mtb. Although histone deacetylases (HDACs), are critical in macrophage differentiations, their roles in macrophage phagocytosis of Mtb remain unexplored. To explore this, I first established human monocyte derived macrophages (MDMs) phagocytosis of red zymosan beads. MDM phagocytosed zymosan beads at 37°C in a dose dependent manner in contrast to suppressed phagocytosis by the MDMs incubated with beads at 4°C or in the presence of cytochalasin D, actin polymerization inhibitor. Incubation of MDMs with 11 zinc-dependent HDAC inhibitors did …

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 …

Generation Of Chimeric Rhinoviruses Presenting Sars-Cov-2 Broadly Neutralizing Epitopes And Their Antigenicity Characterization, Danish Ansari

Biotechnology Theses

The global COVID pandemic is not yet fully under control as there were over 21 million new cases of SARS-CoV-2 infections and over 50,000 deaths globally as of January of 2022. A heavily mutated variant of concern, Omicron is responsible for most of these cases which demands an urgency for a new vaccine. NIH reports over 180 vaccine candidates that use various strategies currently in development. However, a recurring concern with these vaccines is that the continuous viral mutations decrease the efficacy of vaccines. Therefore, we proposed to construct a human rhinovirus (HRV) based chimeric virus containing highly conserved, broadly …

Examination Of The Time Delayed Induction Between Prior Encapsulation Of Catalytic Enzymes In P22 Virus-Like Particles, Andrea Hernandez Irias

Chemistry Theses

Protein cages found in nature have the ability to protect and develop new nanomaterials in order to enhance catalytic reactions. This is due to the ability of these organelle structures to mimic protein-based organelles such as Virus-Like Particles (VLPs). VLPs have the ability to not only resemble virus protein structures but to encapsulate enzymes while retaining their activity. This research examines the in vitro encapsulation withing the bacteriophage P22 derived VLP, and show that some enzymes may require a delay in encapsulation to allowed proper folding

and maturation before they can be encapsulated inside P22 as fully active enzymes. Exploring …

Isolation And Characterization Of Α-Hcd7 Scfv Expressing Exosomes And Synthesis Of Hybrid Exosome-Liposome For Analyses, Sahil Patel

Biotechnology Theses

The goal of this thesis project is to isolate and characterize exosomes that express anti-human CD7 (α-hCD7) single chain variable fragment (scFv), which will be further utilized for T celltargeted gene editing. To reach this goal, exosomes from HEK 293 cells and Jurkat cells were isolated and characterized with different methodologies. To obtain α-hCD7 scFv-expressing exosomes, the HEK 293 cells were subjected to transfection of custom-plasmid pDisplay-αhCD7 scFv, and the expression of α-hCD7 scFv on the HEK 293-derived exosomes were then confirmed by immunogold staining, western blotting, and flow cytometry. To transfect 293 cells, calcium phosphate transfection was utilized, and …