Biotechnology Commons^™

Optimization Of L-Citrulline Production In Corynebacterium Glutamicum, And The Effect Of The Human Small Intestine Environment, Simran S. Ghulldu

Honors Undergraduate Theses

L-citrulline is a crucial amino acid involved in nitric oxide production, which supports cardiovascular health, physical performance, and intestinal well-being. Nitric oxide deficiency is linked to conditions like hypertension, highlighting the need for efficient L-citrulline production. This study attempts to enhance L-citrulline synthesis in Corynebacterium glutamicum, a bacterium widely used in industrial amino acid production, as well as test a The hypothesis is that overexpressing carAB, which encodes carbamoyl phosphate synthetase, will increase carbamoyl phosphate levels and boost L-citrulline production. Two C. glutamicum strains were used: a wild type and the CRBG strain, which has mutations that promote …

Investigating The Role Of Prostasin In Angiogenesis: A Chorioallantoic Membrane Model Approach, Zaima Aline

Honors Undergraduate Theses

Prostasin, also known as PRSS8 or CAP1, is a glycosylphosphatidylinositol (GPI)-anchored extracellular serine protease involved in various physiological and pathophysiological processes. Prostasin expression is observed during key phases of trophoblast invasion and placental development, critical steps in embryo implantation in Rhesus monkeys. This suggests a role for prostasin in extracellular matrix remodeling and tissue invasion, both essential for a healthy pregnancy. Moreover, prostasin appears vital for development as studies in mice show that a germline knockout of the gene leads to significant placental defects and eventual embryonic death, highlighting its importance in vascular and structural development within the placenta.

Building …

Unravelling The Role Of Two-Component Systems In Mycobacterium Abscessus, Andrea C. Molero Perez

Honors Undergraduate Theses

Mycobacterium abscessus (Mab) is an emerging pathogen within the nontuberculous bacteria (NTM) which poses a threat due to its complex multi-drug resistance and opportunistic nature. It is predicted that only 45.6% of patients will have successful treatment, with immunocompromised and cystic fibrosis patients being the most threatened. Moreover, investigating new treatment approaches is key to responding to this public health issue. Two-component systems (TCS) are regulatory pathways that play a pivotal role in bacterial survival and virulence. These systems are composed of a sensor histidine kinase (HK) and cognate response regulator (RR) protein that senses environmental cues and triggers specific …

Developing A Novel Gold Nanoparticle-Based Colorimetric Assay For The Detection Of Cytomegalovirus (Cmv) In Pediatric-Derived Urine Specimens, Sonam Gupta

Graduate Thesis and Dissertation 2023-2024

Cytomegalovirus (CMV) is a member of the Herpesviridae family and is known to infect people of all ages. In most cases, CMV infection is asymptomatic, and the virus is cleared from the host without showing any significant symptoms. However, 1 out of 200 babies are born with congenital CMV infection, which affects multiple organs, including the brain, liver, spleen, lung, and inner ear. One long-term health problem in 1 out 5 babies born with congenital CMV infection is hearing loss. The progression of CMV-associated hearing loss in the first two years of life may lead to developmental delays in language, …

The Ethics Of Brain-Machine Interfaces, Devon J. Lynn

Honors Undergraduate Theses

Brain Machine Interfaces (BMI) are a rapidly developing technology that raise unique ethical issues that demand review. They have demonstrated impressive restorative potential, particularly for individuals living with epilepsy, and those who are locked in. Although BMIs have the potential to provide significant benefit to millions of users, further advancement of the technology should proceed cautiously, according to the guidelines outlined in this paper. Failure to adhere to ethical guidelines could lead to severe privacy concerns, and would violate moral principles of beneficence, virtue ethics, care ethics, and utilitarianism. Despite the moral risks, BMIs hold promise for reshaping future healthcare …

Examining The Effects Of Artemisia Annua On Neuro2a Cells, Wesley Lim

Honors Undergraduate Theses

Artemisia annua L. is a well-known medicinal herb used in Traditional Chinese Medicine for many centuries and contains artemisinin, which exhibits antimalarial properties [1] and is a potential treatment for SARS-CoV-2 [2]. However, the neurological effect of A. annua and its compounds are poorly understood. We hypothesize that A. annua extracts and artemisinin will provide protection against oxidative stress in mouse neuronal Neuro2A cells. Superoxide dismutase 1 (SOD1) is an anti-oxidant enzyme that protects against oxidative damage [3]. Growth Arrest and DNA Damage-inducible 45 alpha (Gadd45a) is involved in DNA repair and is a common biomarker for DNA damage[4]. SOD1 …

Hybridization Kinetics Of Four-Way Junctions Localized On A Dna Scaffold, Katherine N. Taylor

Honors Undergraduate Theses

DNA computing is an ever-growing field with scientists trying to design structures that optimize logic gate communication to develop fast, biologically compatible, computational structures. We hypothesize that by using the principles of DNA computing, it is possible to design a DNA tile capable of studying localized DNA hybridization that can differentiate between oligonucleotides of different structural conformations. This includes synthetically manipulating DNA into a nanostructure that can perform Boolean logic functions to calculate the different rates of hybridization. To test our hypothesis, we designed a DNA Tile that incorporated a 4WJ using YES logic. Linear and hairpin single-stranded (ss) DNA …

A Tool To Automate Neuropathological Assessment In Huntington Disease Mouse Models, Samuel A. Moldenhauer

Honors Undergraduate Theses

As our life expectancy continues to rise, so does the prevalence of neurodegenerative diseases, such as Huntington disease (HD). Neurodegeneration leads to progressive regional brain atrophy, typically initiating prior to symptom onset. With the advancements of medical treatments designed to target neurodegeneration, researchers measure their impact on atrophy in animal models to assess their effectiveness. This is important because treatments designed to combat neuropathology are more likely to modify the disease itself, per contra to treatments designed to mask or treat symptoms. One method of brain region size quantification is magnetic resonance imaging (MRI), which while accurate, is prohibitively expensive. …

Preclinical Assessment Of Psychoplastogens For Treatment Of Psychiatric Symptoms And Neuroplasticity Deficits In Huntington Disease, Nicholas Skiados

Electronic Theses and Dissertations, 2020-2023

Huntington disease (HD) is a neurodegenerative disorder caused by a CAG trinucleotide repeat expansion in the huntingtin (HTT) gene. HD manifests as a triad of psychiatric, cognitive, and motor symptoms, the latter of which defines symptomatic onset of the disease. Psychiatric symptoms including aggression and depression often emerge decades prior to motor symptom onset. Psychiatric disorders are closely linked with dysregulated neural plasticity, the ability of the brain to form new synaptic connections. Neural plasticity is impaired in HD, potentially playing a role in psychiatric symptoms. However, the association between plasticity and psychopathology in HD has yet to be thoroughly …

Extracellular Vesicle-Associated Biomolecules As Potential Biomarkers For Alzheimer's Disease Diagnosis, Lina Bedoya Martinez

Electronic Theses and Dissertations, 2020-2023

Alzheimer's disease (AD) involves progressive neurodegeneration leading to the loss of normal neuronal function. Extracellular accumulation of amyloid-beta (Aß), through the abnormal cleavage of the amyloid precursor protein (APP) by ß- and γ-secretases, is one of the hallmarks of AD. Current research focuses on finding potential candidates for biomarkers and techniques with improved sensitivity for early disease detection. Extracellular vesicles (EVs) found in body fluids are a source of biomarkers for AD diagnosis. EVs transport pathologically significant biomolecules, like nucleic acids and proteins, across the blood-brain barrier, mediating local and distant cell-to-cell communication. Therefore, this study evaluated EV-associated DNA and …

Design And Development Of Medical Devices For Multifaceted Applications, Madisyn Messmore

Honors Undergraduate Theses

The fields of biotechnology and biomedical sciences are rapidly evolving and involve the constant growth of knowledge. As a consequence, engineering design has to also remain at the cutting edge in order to not inhibit the growth of these fields. This study focuses on engineering design and analysis as it pertains to the field of biotechnology, at every step of the engineering process. More specifically, how the engineering design and analysis approach can assist in solving medical problems relating to bone diseases and biomaterials. The first part of the study focuses on a project to design and manufacture a novel …

Intrahost Diversity Of Staphylococcus Aureus In Carriage Among Native Americans, Julia Webb

Electronic Theses and Dissertations, 2020-2023

Staphylococcus aureus (SA) is an opportunistic pathogen and a commensal member of the human microbiota that frequently colonizes the upper respiratory tract and skin. While SA can cause disease ranging from minor skin infections to life-threatening septicemia, it can also be carried asymptomatically. As carriage is the most significant risk factor for SA disease, surveillance is important for the prevention of outbreaks in vulnerable communities. Native Americans have a greater risk of infectious disease than the general US population, and Native Americans in the Southwestern United States have been shown to experience high rates of invasive SA disease. Here we …

The Role Of Ms-818 In Altering Age-Related Characteristics Of An In Vitro Model Of Senescence In Neural Stem Cells, Sandeep Sreerama

Honors Undergraduate Theses

Aging of the brain is the leading risk factor for neurodegenerative diseases and brain cancers and has deleterious effects on brain functions. It follows that attempts to reverse the aging process may be therapeutically valuable. Neural stem cells (NSC) have been shown to play a critical role in maintaining brain functions, and their number is severely decreased with age. The development of senescence-like characteristics and declining functions in NSCs have been proposed to be responsible for brain aging and tumorigenesis. MS-818 is a pyrrolopyrimidine that has been shown to increase the NSC population and reverse the decline of behavioral function …

Competence Transcriptome Changes In Streptococcus Pneumoniae, Smitha Shambhu

Electronic Theses and Dissertations, 2020-2023

Streptococcus pneumoniae is a human commensal and the causative agent of pneumococcal disease. Pneumococci are naturally competent – able to uptake exogenous DNA from the environment and incorporate it into their genome through homologous and non-homologous recombination. Recombination has significantly shaped the evolutionary history of S. pneumoniae, as it allows pneumococci to rapidly adapt to interventions such as antibiotic therapy or vaccine introduction. Recombination frequencies vary considerably across pneumococcal populations; yet the underlying mechanisms for these variations are not well understood. Entry and exit into competence, a state in which the cell can uptake DNA, is tightly regulated through transcriptional …

The Effect Of A Senolytic Drug On Messenger Rna And Micro Rna Expression In Perigonadal Adipose Tissue Of Young And Old Mice, Catalina Hernandez

Honors Undergraduate Theses

As we face an expanding elderly population and increased age-related chronic diseases, we must seek solutions that address interconnected problems. Dasatinib plus quercetin (D+Q), a novel combination drug, is a proposed solution to improve the health and longevity of aged individuals with minimal side effects, exceptional efficacy, and maximum convenience. In this experiment, intermittent D+Q treatment was completed on young and old mice. RNA was extracted from perigonadal adipose tissue and cDNA libraries of mRNA and miRNA were made. Real-time quantitative polymerase chain reaction was used to assess relative gene expression for mRNA and miRNA pertaining to cell senescence, inflammation, …

Identification Of Novel Antimalarials From Natural Product Inspired Libraries, David Perry

Electronic Theses and Dissertations, 2020-2023

To identify novel antimalarials from unexplored areas of chemical space, we have utilized two approaches: (a) using Torrey Pines Institute for Molecular Studies (TPIMS) proprietary high-density combinatorial libraries containing a large number of small molecule compounds residing largely in the underexplored areas of chemical space; and, (b) using novel, complexity-to-diversity/ring distortion of available natural products. The first approach involved the screening of more than 30 million compounds derived from 81 small molecule libraries, built on 81 distinct scaffolds. From this, we identified the pyrrolidine bis-cyclic guanidine library (TPI-1955) to be one of the most active and selective for antiplasmodial activity. …

Zika Virus-Induced Lysis Of Cervical Cancer Cells, Harini Krishnapura

Electronic Theses and Dissertations

Cervical cancer is the fourth most frequent cancer in women with an estimated 570,000 new cases globally in 2018. Treatment of advanced cervical cancer is often unsuccessful leading to high cancer-related mortality rates, especially in under-resourced countries. Recently, a possible role for the cell surface glycoprotein CD24 in host cell specificity of Zika virus was reported. As an extension of this work, Zika viruses have been proposed as oncolytic therapy for the treatment of neuroblastoma and other CD24 positive tumors. To determine the permissiveness of cervical cancer cells to Zika virus infection and its association with CD24, we assessed cytopathic …

Autologous Skin Cell Spray-Transplantation As An Innovative Alternative To Autologous Split- Thickness Skin Grafts For Deep Partial Thickness Burn Wounds: An Integrative Literature Review, Alexandria M. Beaudet

Honors Undergraduate Theses

Burn wounds tend to be a critical problem with a complicated healing process. Although advancements have been made and the treatment of burn wounds has improved significantly, the healing process for deep-partial thickness burn wounds remains problematic. The purpose of this thesis is to review the available literature on an innovate biotechnology, autologous skin cell-spray transplantation, to more effectively treat burn wounds and potentially other injuries in the future. This study was conducted by critically researching and comparing (N=7) peer-reviewed research articles focusing not only on burn wounds using traditional treatments, but also the treatment of burn wounds using revolutionary …

Metabolic Effects Of 17a-Estradiol Are Growth Hormone Independent And Sex Specific, Silvana Sidhom

Electronic Theses and Dissertations

Aging is a major risk factor for metabolic syndromes and type two diabetes. With growing elderly populations worldwide and increasing incidence of age-related diseases there is a great need to develop pharmacological interventions that would delay aging and protect from age-related diseases. 17-alpha estradiol (17α-E2) is an epimer of the primary female sex hormone estradiol and has been shown to extend lifespan and downregulate markers of age-related metabolic dysfunction in male mice. Because 17α-E2 does not induce feminization in males it holds potential as a novel therapeutic in humans for age-related metabolic dysfunction. Importantly, we have previously shown that 17α-E2 …

Identification And Functional Characterization Of A Long Non-Coding Rna Associated With Prostate Cancer, Md Faqrul Hasan

Electronic Theses and Dissertations

Prostate cancer is the most common cancer in men in the western world. Although early stage prostate cancer is treatable late stage, more specifically, metastatic and drug resistant prostate cancers are mostly incurable. The failure of current treatments obligates the research community to explore novel areas in prostate cancer biology and find better therapeutic targets. Emerging evidences show that non-coding RNAs specifically long non-coding RNAs (lncRNAs) play regulatory roles in various cellular processes and are frequently dysregulated in cancer including prostate cancer. These aberrantly expressed lncRNAs mostly with unexplored genetic information may drive cancer progression. Previous studies done in our …

Unraveling The Role Of Phenylethanolamine N-Methyltransferase (Pnmt+) Cells In-Vivo, Sanjana Manja

Electronic Theses and Dissertations

Phenylethanolamine N-methyltransferase (Pnmt) is the enzyme that N-methylates norepinephrine to produce the stress hormone/neurotransmitter, epinephrine, which is abundantly expressed in adrenal glands. Developmental studies have also identified Pnmt expression in the embryonic heart and several areas of the brain, including brainstem, cerebellum, and hypothalamus. Thus, we hypothesize that selective ablation of Pnmt+ cells will have detrimental effects on cardiovascular, neuromuscular, and metabolic processes. To uncover the importance of Pnmt+ cells in vivo, we generated a novel Diphtheria Toxin A (DTA) suicide model (Pnmt+/Cre; R26+/DTA) to selectively ablate Pnmt-expressing (Pnmt+) cells in mice. Appearing normal at birth, Pnmt-Cre/DTA mice began to …

Bone Morphogenetic Protein 7 Inhibits Pyroptotic Cell Death In Vascular Smooth Muscle Cells Of Atherosclerotic Apolipoprotein E -/- Mice, Kaley Garner

Electronic Theses and Dissertations

Atherosclerosis (ATH) is an inflammation-mediated disease in which cell death underlies the formation of lesions along the intima layer of vascular walls resulting in vessel narrowing, decreased blood flow, and increased risk of lesion rupture leading to myocardial infarction and stroke. The current study was undertaken to investigate whether inflammation in ATH can induce pyroptosis in vascular smooth muscle cells (SMC's). We therefore hypothesized that pyroptosis occurs and is inhibited by bone morphogenetic protein 7 (BMP7). We examined SMC pyroptosis at acute (D5) and midstage (D28) following disturbed flow-induced hemodynamic injury to the vascular wall using our partial left carotid …

Selenium Vs. Sulfur: Investigating The Substrate Specificity Of A Selenocysteine Lyase, Michael Johnstone

Electronic Theses and Dissertations

Selenium is a vital micronutrient in many organisms. While traces are required for survival, excess amounts are toxic; thus, selenium can be regarded as a biological "double-edged sword". Selenium is chemically similar to the essential element sulfur, but curiously, evolution has selected the former over the latter for a subset of oxidoreductases. Enzymes involved in sulfur metabolism are less discriminate in terms of preventing selenium incorporation; however, its specific incorporation into selenoproteins reveals a highly discriminate process that is not completely understood. In this work, we add knowledge to the mechanism for selenium-over-sulfur specificity in hopes of further understanding the …

Determining Differential Effects Of Interleukin-2 On Innate And Adaptive Immune Cells In Lymphoid Organs And The Gastrointestinal Tract, Ayushi Singh

Electronic Theses and Dissertations

Interleukin-2 (IL-2) is a pleiotropic cytokine demonstrated to be effective in treating cancer. However, the clinical use of IL-2 can be associated with severe side effects including gastrointestinal toxicity (GT). Similar GT symptoms are observed in inflammatory diseases such as CD (CD). Interestingly mounting evidence indicates a role for IL-2 in CD, but the underlying mechanisms are unknown. Indeed, studies on the in-vivo activities of IL-2 have mostly focused on secondary lymphoid organs and immune cells associated with them. Very few studies have addressed how IL-2 signals impact populations of immune cells in the gut. Here, we aim to identify …

Atp Induced Molecular Disassembly Of Cytolethal Distending Toxin's B/C Heterodimer, George Huhn

Electronic Theses and Dissertations

Cytolethal distending toxin (CDT) is a virulence factor produced by many Gram-negative bacteria, including Haemophilus ducreyi. This fastidious pathogen is the causative agent of genital cancroid. CDT is a heterotrimeric toxin with an AB2 structure consisting of a cell-binding "B" domain (CdtA + CdtC) and a catalytic "A" domain (CdtB) that has DNase activity. This toxin assembles in the bacterial periplasm that lacks ATP and is secreted into the extracellular environment. After cell binding, CDT is internalized by endocytosis and travels through the endosomes and Golgi before arriving in the endoplasmic reticulum (ER). CdtA is lost from the holotoxin before …

Biochemical Characterization Of Rv2633c From Mycobacterium Tuberculosis And The Effects Of Mutagenesis On Iron Binding, Kyle Strickland

Electronic Theses and Dissertations

Mycobacterium tuberculosis (Mtb) is a pathogenic bacterium that is the causative agent of the disease Tuberculosis (TB). TB kills an estimated 1.8 million people annually and roughly one third of the world's population carries Mtb in a dormant state. Drug resistant Mtb strains are on the rise, thus a new method of combating this disease is paramount. Mtb survival inside of macrophages requires overcoming various stressors such as; iron restriction, reactive oxygen species, and hypoxic conditions. Mtb employs the use of catalases, nitric oxide reductase, superoxide dismutase, and siderophores to aid in survival. These functions have also been found in …

Role Of Single Nucleotide Polymorphisms (Snps) In Ptpn2/22 And Mycobacterium Avium Subspecies Paratuberculosis (Map) In Rheumatoid Arthritis And Crohn's Disease, Robert Sharp

Electronic Theses and Dissertations

Both genetic pre-disposition and potential environmental triggers are shared between Rheumatoid arthritis (RA) and Crohn's disease (CD). We hypothesized that single nucleotide polymorphisms (SNPs) in the negative T-cell regulators Protein Tyrosine Phosphatase Nonreceptor type 2 and 22 (PTPN2/22) lead to a dysregulated immune response as seen in RA and CD. To test the hypothesis, peripheral leukocytes samples from 204 consented subjects were TaqMan genotyped for 9 SNPs in PTPN2/22. The SNPs effect on PTPN2/22 and IFN-y expression was determined using RT-PCR. Blood samples were analyzed for the Mycobacterium avium subspecies paratuberculosis (MAP) IS900 gene by nPCR. T-cell proliferation and response …

Pg-Vtv Biogenesis Requires Atp To Facilitate Phosphorylation Of Syntaxin 17, Anika Saxena

Electronic Theses and Dissertations

The uptake of cytotoxic free fatty acids (FFA) and their conversion to physiologically expedient triglycerides (TAG) which are later on assimilated to very low density lipoproteins (VLDL) is of utmost value among the multifarious tasks performed by the liver. Inflated concentration of VLDL in the blood stream directly correlates with the reinforcement of atherosclerosis. VLDL is synthesized in the hepatic endoplasmic reticulum (ER) and transported to the Golgi where it encounters several alterations. It is then enclosed in distinct post-Golgi VLDL transport vesicles (PG-VTVs) and released into the blood. The data generated in our lab has proved the requirement of …

Deciphering The Role Of Adrenergic Hormones In Embryonic Cardiac Calcium Signaling And Metabolism, Jessica Peoples

Electronic Theses and Dissertations

The adrenergic hormones norepinephrine (NE) and epinephrine (EPI) are critical regulators of mammalian cardiovascular physiology. NE and EPI mediate stress responses to enhance cardiovascular function, however dysregulation of adrenergic signaling leads to heart failure, congenital heart malformations, and sudden cardiac death. Adrenergic hormone-expressing cells were found in the early embryonic heart, and NE has been determined essential for embryonic cardiac development. Despite extensive work in adults, the regulatory roles and adrenergic targets of these hormones during embryonic cardiac development have not yet been fully determined. Prior transcriptomic studies from our lab showed that expression of signal transduction and metabolic genes …

Characterization Of Neural Cells Derived From Reelin-Deficient Schizophrenic Patient Ips Cells, Nicole Roberts

Electronic Theses and Dissertations

Reelin is a large, extracellular glycoprotein that binds to several membrane receptors on neural stem cells (HNSCs), neural progenitor cells (NPCs), and neuroblasts of mammals to direct their migration. Previously, our lab established the presence of Reelin increased migration of wild-type fetal-derived HNSC's, both in vitro and in vivo. In addition, we demonstrated that Reelin protein treatment also increases the formation of radial glia via Notch-1 signaling, in vitro. Radial glia are precursors to NPCs, as well as a scaffold for neuroblast migration during cortical lamination. Reelin has long been associated with Schizophrenia (SZ). Because post-mortem brains are limited to …