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Full-Text Articles in Life Sciences

Investigating The Role Of Lbh During Early Embryonic Development In Xenopus Laevis, Emma Weir Oct 2019

Investigating The Role Of Lbh During Early Embryonic Development In Xenopus Laevis, Emma Weir

Masters Theses

LBH is a highly conserved protein whose role during vertebrate development is relatively under-studied. In collaboration with the Albertson lab, our lab has previously shown that it is necessary for cranial neural crest cell migration in the zebrafish and in Xenopus laevis. The molecular mechanisms through which it acts are not well understood.

In Xenopus, LBH is a maternally deposited protein. As such, studying its role in early development has not been feasible through the morpholino-mediated knockdown techniques that prevent translation of target genes. Recently, a technique for degrading endogenous proteins was developed, called Trim-Away. This was developed in mammalian ...


Protein-Nanoparticle Co-Engineering: Self-Assembly, Intracellular Protein Delivery, And Crispr/Cas9-Based Gene Editing, Rubul Mout Nov 2017

Protein-Nanoparticle Co-Engineering: Self-Assembly, Intracellular Protein Delivery, And Crispr/Cas9-Based Gene Editing, Rubul Mout

Doctoral Dissertations

Direct cytoplasmic delivery of gene editing nucleases such CRISPR/Cas9 systems and therapeutic proteins provides enormous opportunities in curing human genetic diseases, and assist research in basic cell biology. One approach to attain such a goal is through engineering nanotechnological tools to mimic naturally existing intra- and extracellular protein delivery/transport systems. Nature builds transport systems for proteins and other biomolecules through evolution-derived sophisticated molecular engineering. Inspired by such natural assemblies, I employed molecular engineering approaches to fabricate self-assembled nanostructures to use as intracellular protein delivery tools. Briefly, proteins and gold nanoparticles were co-engineered to carry complementary electrostatic recognition elements ...


Pore Forming Protein Assembly And The Use In Nanopore Sensing: A Study On E. Coli Proteins Clya And Ompg, Monifa Fahie Nov 2017

Pore Forming Protein Assembly And The Use In Nanopore Sensing: A Study On E. Coli Proteins Clya And Ompg, Monifa Fahie

Doctoral Dissertations

Pore forming proteins are typically the proteins that form channels in membranes. They have several roles ranging from molecule transport to triggering the death of a cell. This work focuses on two E. coli pore forming proteins that have vastly differing roles in nature. Outer membrane protein G (OmpG) is an innocuous β-barrel porin while Cytolysin A (ClyA) is an α-helical pore forming toxin. For OmpG we probed its potential to be a nanopore sensor for protein detection and quantification. A small high affinity ligand, biotin, was covalently attached to loop 6 of OmpG and used to capture biotin-binding proteins ...


Microbial Dynamics And Design Considerations For Decentralized Microbial Fuel Cell Applications, Cynthia Castro Nov 2017

Microbial Dynamics And Design Considerations For Decentralized Microbial Fuel Cell Applications, Cynthia Castro

Doctoral Dissertations

The purpose of this dissertation was to assess the practicality of using microbial fuel cells (MFCs) as alternative sanitation systems for wastewater treatment and energy recovery, focusing on identifying key design considerations for treating high strength wastewater and managing alternative metabolic pathways.

We evaluated the energetic outputs of a lab-based pilot MFC designed to treat complex organics present in both synthetic feces and municipal wastewater. The pilot MFC produced two energetic products, methane and electricity, when treating two types of complex wastewaters. The energetic products associated with anode respiration and methanogenesis were simultaneously observed and yielded a combined energy ouput ...


Robust Biosensors For Healthcare Applications: From High-Content Screening To Point-Of-Care Testing, Ngoc D. B. Le Nov 2017

Robust Biosensors For Healthcare Applications: From High-Content Screening To Point-Of-Care Testing, Ngoc D. B. Le

Doctoral Dissertations

Efficient detection of proteins, mammalian cells, microorganisms and other biological systems in complex mixture is essential in disease diagnosis and environmental health. Therefore, technological platforms that provide sensors of high sensitivity, selectivity and stability are greatly desired. Recently, the ‘chemical-nose’ sensing approach has proved to be an effective strategy for profiling bio-relevant targets in complex mixtures. Detecting analytes in complex mixture is a challenge that conventional specificity-based sensors are still trying to solve due to the requirement of prior knowledge of the analyte, which is unknown in many cases. This thesis focuses on how to develop simple and robust ...


Novel Advancements For Improving Sprout Safety, Kyle S. Landry Jul 2016

Novel Advancements For Improving Sprout Safety, Kyle S. Landry

Doctoral Dissertations

All varieties of bean sprouts (mung bean, alfalfa, broccoli, and radish) are classified as a “super-food” and are common staples for health conscious consumers. Along with the proposed health benefits, there is also an inherent risk of foodborne illness. When sprouts are cooked, there is little risk of illness. The purpose of this dissertation was to explore novel techniques to minimize or prevent the incidence of foodborne illness associated with the consumption of sprouts. Three areas were investigated: 1) the use of a biocontrol organism, 2) the use of a novel spontaneous carvacrol nanoemulsion, and 3) the influence of the ...


Promoting Extracellular Matrix Crosslinking In Synthetic Hydrogels, Marcos M. Manganare Nov 2015

Promoting Extracellular Matrix Crosslinking In Synthetic Hydrogels, Marcos M. Manganare

Masters Theses

The extracellular matrix (ECM) provides mechanical and biochemical support to tissues and cells. It is crucial for cell attachment, differentiation, and migration, as well as for ailment-associated processes such as angiogenesis, metastases and cancer development. An approach to study these phenomena is through emulation of the ECM by synthetic gels constructed of natural polymers, such as collagen and fibronectin, or simple but tunable materials such as poly(ethylene glycol) (PEG) crosslinked with short peptide sequences susceptible to digestion by metalloproteases and cell-binding domains. Our lab uses PEG gels to study cell behavior in three dimensions (3D). Although this system fosters ...


Studying Nanoparticle/Cell And Nanoparticle/Biosurface Interaction With Mass Spectrometry, Singyuk Hou Nov 2015

Studying Nanoparticle/Cell And Nanoparticle/Biosurface Interaction With Mass Spectrometry, Singyuk Hou

Masters Theses

Nanoparticles (NPs) have been used widely in various fields ranging from biomedical applications to life science due to their highly tunable properties. It is essential to understanding how NPs interact with biological systems of interest, therefore, analytical platforms to efficiently track NPs from cell to animal level are essential. In this thesis, laser desorption ionization mass spectrometry (LDI-MS) and inductively-coupled plasma mass spectrometry (ICP-MS) has been developed and applied to quantify NP/cell and NP/biological surface interactions. These two methods provide fast, label-free and quantitative analysis. New capability of LDI-MS to differentiate cell surface-bound and internalized NPs were established ...


Design And Synthesis Of Polymeric Nanoparticles For Drug And Protein Delivery, Judy A. Ventura Aug 2015

Design And Synthesis Of Polymeric Nanoparticles For Drug And Protein Delivery, Judy A. Ventura

Doctoral Dissertations

Nanoparticles are emerging as carriers in biological applications due to advances in their preparation, size control, surface modification and encapsulation capabilities. In addition, nanomaterials improve bioavailability by enhancing aqueous solubility of the guest molecule and increasing resistance time in the body. However, the delivery of guest molecules is still challenging due to the intrinsic characteristics of the guest molecule including large size and propensity to denature or degradation in the case of biomolecules and the encapsulation stability of the small guest molecules.

Our group recently reported the preparation of self-cross-linked polymeric nanogels possessing surface functionalization capabilities. In this dissertation we ...


Engineering Novel Detection And Treatment Strategies For Bacterial Therapy Of Cancer, Jan T. Panteli Aug 2015

Engineering Novel Detection And Treatment Strategies For Bacterial Therapy Of Cancer, Jan T. Panteli

Doctoral Dissertations

Finding and treating cancer is difficult due to limited sensitivity and specificity of current detection and treatment strategies. Many chemotherapeutic drugs are small molecules that are limited by diffusion, making it difficult to reach cancer sites requiring high doses that lead to systemic toxicity and off-target effects. Tomographic detection techniques, like PET, MRI and CT, are good at identifying macroscopic lesions in the body but are limited in their ability to detect microscopic lesions. Biomarker detection strategies are extremely sensitive and able to identify ng/ml concentrations of protein, but are poor at discriminating between healthy and disease state levels ...


Transferrin-Based Therapeutics And Analytical Methods To Characterize Them, Son N. Nguyen Aug 2015

Transferrin-Based Therapeutics And Analytical Methods To Characterize Them, Son N. Nguyen

Doctoral Dissertations

Modern development of medicine requires detailed characterization by state-of-the art analytical techniques that can be used to analyze covalent structure, conformations and protein-receptor interaction to quantitatively measure biodistribution of protein therapeutics. Mass spectrometry has already become an indispensable tool facilitating all stages of protein drug development. Particularly, this work has demonstrated the tremendous potential of electrospray ionization (ESI) mass spectrometry (MS) in this arena by providing invaluable information beyond mass measurement that can be used to optimize protein drug conjugate structures during early stages of development, and to further catalyze drug design efforts. Additionally, a new sensitive and selective method ...


Rnai Mediated Silencing Of Cell Wall Invertase Inhibitors To Increase Sucrose Allocation To Sink Tissues In Transgenic Camelina Sativa Engineered With A Carbon Concentrating Mechanism, Joshua Zuber Jul 2015

Rnai Mediated Silencing Of Cell Wall Invertase Inhibitors To Increase Sucrose Allocation To Sink Tissues In Transgenic Camelina Sativa Engineered With A Carbon Concentrating Mechanism, Joshua Zuber

Masters Theses

Plant invertases are a class of proteins that have enzymatic function in cleaving sucrose to fructose and glucose. Cell wall invertase, located on the exterior of the cell wall of plant cells, plays a key role in the unloading of sucrose from the apoplast to the sink tissues. Cell wall invertase interacts with an inhibitor, cell wall invertase inhibitor, post-transcriptionally to regulate its activity. The inhibitor is constitutively expressed in pollen development, early developing seeds, and senescing leaves: indicative of sucrose allocation being a limiting factor at these stages of development. We introduced algal bicarbonate transporters LCIA/CCP1 to Camelina ...


Agricultural Waste Remediation And H2 Production By Hyperthermophilic Heterotrophs: Bioinformatics, Taxonomy, And Physiology, Sarah A. Hensley Mar 2015

Agricultural Waste Remediation And H2 Production By Hyperthermophilic Heterotrophs: Bioinformatics, Taxonomy, And Physiology, Sarah A. Hensley

Doctoral Dissertations

In the United States, nearly 35 million tons of food waste are delivered to landfills and wastewater treatment plants for disposal each year. This places a burden on landfills and the environment through the introduction of foreign chemicals. Utilizing hyperthermophilic heterotrophs for waste treatment in a consolidated bioprocess could potentially remediate these organics, increase reaction rates, kill pathogens, produce a bioenergy product (H2), and lower costs relative to mesophilic waste treatment options. However, it is unknown how various agricultural waste streams will affect hyperthermophilic growth and metabolite production. We investigated Thermococcales species for this purpose. Specifically, we chose Pyrococcus ...


Understanding The Transcriptional Regulation Of Secondary Cell Wall Biosynthesis In The Model Grass Brachypodium Distachyon, Pubudu Handakumbura Nov 2014

Understanding The Transcriptional Regulation Of Secondary Cell Wall Biosynthesis In The Model Grass Brachypodium Distachyon, Pubudu Handakumbura

Doctoral Dissertations

Secondary cell wall synthesis occurs in specialized cell types following completion of cell enlargement. By virtue of mechanical strength provided by a wall thickened with cellulose, hemicelluloses, and lignin, these cells can function as water-conducting vessels and provide structural support. Several transcription factor families regulate genes encoding wall synthesis enzymes. Certain NAC and MYB proteins directly bind upstream of structural genes and other transcription factors. The most detailed model of this regulatory network is established predominantly for a eudicot, Arabidopsis thaliana. In grasses, both the patterning and the composition of secondary cell walls are distinct from that of eudicots. These ...


Engineering Camelina Sativa For Biofuel Production Via Increasing Oil Yield And Tolerance To Abiotic Stresses, Kenny Ablordeppey Nov 2014

Engineering Camelina Sativa For Biofuel Production Via Increasing Oil Yield And Tolerance To Abiotic Stresses, Kenny Ablordeppey

Masters Theses

In an effort to engineer Camelina sativa for enhanced oil yield and tolerance to abiotic stresses, we have cloned and overexpressed Camelina γ-Glutamyl Cyclotransferase (GGCT2;1); a gene involved in oxidative stress tolerance via glutathione homeostasis and Wrinkled 1; a transcription factor that regulates genes involved in fatty acid biosynthesis to increase triacylglycerol (TAG) accumulation in seeds. The GGCT gene family in Camelina consists of three genes-GGCT1, GGCT2;1 and GGCT2;2. Camelina GGCT genes showed differential expression under oxidative stress caused as a result of exposure to various abiotic stresses. The GGCT2;1 gene, which showed strong up-regulation ...


Cd4 Silencing In Thymocytes Is Opposed By The Enforced Association Of P300 Hat, Hdac1 Or Suv39h1 With Runx Transcription Factors, Christyne Ayne Kane Apr 2014

Cd4 Silencing In Thymocytes Is Opposed By The Enforced Association Of P300 Hat, Hdac1 Or Suv39h1 With Runx Transcription Factors, Christyne Ayne Kane

Doctoral Dissertations

The transcription factors Runx1 and Runx3 are required for permanent silencing of CD4 in maturing CD8+ thymocytes. Runx binding to consensus sites within the CD4 silencer region is required for CD4 silencing post-positive selection. The Runx nuclear matrix targeting sequence (NMTS) is required for CD4 silencing and is implicated in binding to the histone acetyltransferase (HAT) p300, histone deacetylases (HDAC), and histone methyltransferase (HMT) SUV39H1 proteins. Epigenetic modifications of chromatin or post-translational modifications of Runx itself as a result of Runx association with these enzymes may be important for establishment of long-term CD4 silencing. In this study, we show that ...


The Role Of Notch In Th17 Differentiation, Reem Suleiman Sep 2013

The Role Of Notch In Th17 Differentiation, Reem Suleiman

Open Access Dissertations

Th17 cells are pro-inflammatory cells that are characterized by the production of their signature cytokine, IL-17. Although they are thought to have arisen to protect against extracellular bacteria and fungi they have been shown to mediate autoimmune diseases such as EAE and psoarisis. Notch protein is a cell-surface receptor that has been widely conserved among species. It plays an essential role in determining multiple cell fates. More recently, it has been implicated in regulating peripheral CD4+ T-cell responses. In these studies, we report that blockade of Notch signaling significantly down-regulates the production of IL-17 and associated cytokines in both mouse ...


Molecular Mechanisms Regulating Complement Receptor 3-Mediated Phagocytosis Of Borrelia Burgdorferi, Kelly Lynn Hawley Sep 2012

Molecular Mechanisms Regulating Complement Receptor 3-Mediated Phagocytosis Of Borrelia Burgdorferi, Kelly Lynn Hawley

Open Access Dissertations

The macrophage receptors that mediate phagocytosis of Borrelia burgdorferi, the Lyme disease spirochete, are unknown despite this cell type's importance in promoting pathogen clearance and inflammation-mediated tissue damage. We now demonstrate that the β2 integrin, Complement Receptor 3 (CR3), mediates the phagocytosis of opsonized and non-opsonized spirochetes by murine macrophages and human monocytes. Although, expression of the surface proteins, CspA and OspE, protects B. burgdorferi from complement-mediated phagocytosis, the versatility of CR3 counteracts the ability of B. burgdorferi to interfere with complement activation and complement-derived opsonins, thus minimizing the bacteria's anti-complement strategy. Interaction of the spirochete with the ...


Innate Immune Responses To B. Burgdorferi Mediated By Jnk1 And The Cochaperone, Methylation Controlled Dnaj (Mcj), Hooman Izadi Feb 2011

Innate Immune Responses To B. Burgdorferi Mediated By Jnk1 And The Cochaperone, Methylation Controlled Dnaj (Mcj), Hooman Izadi

Open Access Dissertations

The infections agent of Lyme disease, Borrelia Burgdorferi is a complex microorganism with a highly diverse genome. One of the most remarkable aspects of the B. burgdorferi genome is the large number of sequences encoding predicted or known lipoproteins, including outer-surface proteins. The B. burgdorferi genome encodes no recognizable toxins. Instead, this extracellular pathogen causes pathology by migration through tissues, adhesion to host cells, and evasion of immune clearance. Inflammation elicited by infection with B. burgdorferi depends on the ability of the spirochete to survive in the mammalian host, as well as the immune response that arises upon the interaction ...


Tyrosine Phosphorylation Events In Mouse Sperm Capacitation, Enid Arcelay Sep 2009

Tyrosine Phosphorylation Events In Mouse Sperm Capacitation, Enid Arcelay

Open Access Dissertations

Mammalian sperm are not able to fertilize immediately upon ejaculation; they become fertilization-competent after undergoing changes in the female reproductive tract collectively termed capacitation. Although it has been established that capacitation is associated with an increase in tyrosine phosphorylation, little is known about the role of this event in sperm function. In this work we used a combination of two dimensional gel electrophoresis and mass spectrometry to identify proteins that undergo tyrosine phosphorylation during capacitation. Some of the identified proteins are the mouse orthologues of human sperm proteins known to undergo tyrosine phosphorylation. Among them we identified VDAC, tubulin, PDH ...