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Characterization And Use Of Folate Receptor Isoforms For Targeting Of Epithelial And Myeloid Cells, Sreya Biswas

Theses and Dissertations

ABSTRACT

CHARACTERIZATION AND USE OF FOLATE RECEPTOR ISOFORMS FOR TARGETING OF EPITHELIAL AND MYELOID CELLS

by

Sreya Biswas

The University of Wisconsin-Milwaukee, 2016

Under the Supervision of Professor Douglas A. Steeber

Folate receptor (FR) is a GPI-anchored glycoprotein with high binding affinity for folic acid. FR has two membrane-associated isoforms, α and β, that are overexpressed on epithelial and myeloid tumors, respectively. Normal cells may also exhibit FR expression at very low levels but interestingly, FR-α on normal cells is restricted to the apical surface i.e., away from the blood stream. This differential expression and orientation of the FR-α isoform …

Symmetry And Reconstruction Of Particle Structure From Random Angle Diffraction Patterns, Sandi Wibowo

Theses and Dissertations

The problem of determining the structure of a biomolecule, when all the evidence from experiment consists of individual diffraction patterns from random particle orientations, is the central theoretical problem with an XFEL. One of the methods proposed is a calculation over all measured diffraction patterns of the average angular correlations between pairs of points on the diffraction patterns. It is possible to construct from these a matrix B characterized by angular momentum quantum number l, and whose elements are characterized by radii q and q’ of the resolution shells. If matrix B is considered as dot product of vectors, which …

Molecular And Functional Analysis Of The Pixb Gene In Xenorhabdus Nematophila, John Lucas

Theses and Dissertations

Xenorhabdus nematophila and the entomopathogenic nematode Steinernema carpocapsae form a mutualistic relationship facilitating the infection, death and consumption of an insect host. The infective juvenile (IJ) form of S. carpocapsae invades the insect host through natural openings and proceeds to the hemocoel where exposure to hemolymph stimulates the release of X. nematophila from the anterior vesicle. Excreted X. nematophila releases immunosuppressive compounds and insect toxins into the insect hemolymph that facilitates death of the host. As X. nematophila reaches high cell density it secretes exoenzymes that degrade insect tissues and produces antibiotics that reduce microbial competition. S. carpocapsae utilizes the …

Discovery And Characterization Of A New Group Of Is10 Insertion Sequences, Rachel Marie Kinzelman

Theses and Dissertations

Insertion sequences (ISs) are small mobile genetic elements that can have significant impact on the genotype and phenotype of a host organism. Previous work in this laboratory revealed an insertion sequence that disrupted the luxA gene in Vibrio harveyi strain BCB451, knocking out light production. Phylogenetic analysis of this insertion sequence, dubbed IS451, reveals that it is in the IS10 family, but represents a novel variant that is only 79% identical to other known IS10 sequences. Twelve copies of IS451 were isolated from a genomic library and sequenced, and were found to be essentially identical, but located in dispersed chromosomal …

The Role Of Proteome In Cellular Zn2+ Trafficking And In The Ability Of The Fluorescent Zinc Sensors To Image Intracellular Zn2+, Mohammad Rezaul Karim

Theses and Dissertations

Zinc is an essential biological trace metal used in as many as 3000 Zn-proteins, about 10% of the eukaryotic proteome, as either a structural constituent or a catalytic cofactor. These proteins include the zinc fingers, the most prevalent transcription factors that bind a wide range of gene promoters and thus regulate gene expression. A eukaryotic cell contains several hundred micromolar of Zn2+- almost all of it is bound to specific Zn-proteins. Recently, Zn2+ has been reported to serve as a regulatory signal and a neurotransmitter, suggesting that there also exists a dynamic Zn2+ pool in cells. These findings led to …

Structural And Functional Characterization Of Acetoacetate Decarboxylase-Like Enzymes, Lisa Mueller

Theses and Dissertations

The acetatoacetate decarboxylase-like superfamily (ADCSF) is a largely unexplored group of enzymes that may be a potential source of new biocatalysts. Bioinformatic analysis has grouped these approximately 2000 enzymes into seven different families based on comparison of predicted active site residues. To date, only the prototypical ADCs (Family I) that catalyze the decarboxylation of acetoacetate have been studied. Analysis of gene context suggests that Family V contains predominantly enzymes predicted to be involved in secondary metabolism. On average, these share about 20% sequence identity to the true ADCs. To learn more about the diversity of chemistries performed by members of …

Lateral Replacement Of The Lux Operon In A Vibrio Isolated From The Intestine Of A Coral Reef Fish, Melissa Lee Whyte

Theses and Dissertations

In a screening of bioluminescent bacteria isolated from the intestines of coral reef fish, two strains (designated D6 and M1) were identified that have a luxA gene sequence significantly different from those of other Vibrio species. Phylogenetic analysis of several housekeeping genes, as well as toxR, shows that D6 and M1 branch within a bioluminescent clade (designated the “D1 group,” isolated at the same time and place as D6 and M1) that is a close sister group to Vibrio harveyi. However, whereas the luxA genes of the D1 group are >98% identical to V. harveyi luxA, the luxA genes of …

The Function Of Renalase, Brett Allen Beaupre

Theses and Dissertations

Renalase was originally reported to be an enzyme secreted into the blood by the kidney to lower blood pressure and slow heart rate. Despite multiple reports claiming to confirm this activity in vivo there has been considerable discord in regards to the reaction catalyzed by renalase. The structural topology of renalase resembles that of known flavoprotein oxidases, monooxygenases and demethylases, but the conserved active site residues are unique to renalase. It has been reported that the catalytic function of renalase is to oxidize circulating catecholamines, however in vitro studies have failed to demonstrate a catalytic activity in the presence of …

The Selective Survival Of Escherichia Coli In Freshwater Beach Sand, Natalie Ann Rumball

Theses and Dissertations

The quantification of Escherichia coli or E. coli is the most common method used to detect recent fecal pollution in recreational water, as this species is known for its high abundance in fecal matter and assumed host-associated nature. However, it has been determined that some strains are capable of long-term survival and potential propagation in non-host environments, such as the beach sand. These long-term environmental survivors are host-independent and are not associated with the same health risks as those E. coli from recent fecal pollution. However, they have been shown to impact how water quality is perceived as they are …

Role Of Protein Phosphatase Reg2-Glc7 In The Regulation Of The Yeast Stress Response Kinase, Snf1, Marcin Maziarz

Theses and Dissertations

Kinases of the AMP-activated protein kinase (AMPK) family are conserved in eukaryotes and play central roles in responses to reduced energy availability. AMPK, nicknamed the “fuel gauge” of the cell, monitors cellular energy status via the ratio of AMP to ATP nucleotides. AMPK restores energy homeostasis by reducing energy “spending” and increasing energy “income”. Correspondingly, defects in AMPK signaling have been implicated in diseases including type II diabetes, obesity, and cancer.

In yeast, the AMPK homolog is Snf1 protein kinase. Glucose is the preferred carbon/energy source of yeast, and thus limitation for glucose similarly activates Snf1. Snf1 activation requires phosphorylation …

Protective Effects Of Pigmentation In Pseudomonas Aeruginosa: Insights On Pyomelanin Production And Inhibition By Ntbc, Laura Marie Ketelboeter

Theses and Dissertations

Pyomelanin is an extracellular, reddish-brown pigment produced by several environmental and pathogenic bacteria and fungi. It is derived from the phenylalanine/tyrosine catabolism pathway and is produced when homogentisate (HGA) is secreted from the cell, auto-oxidized, and self-polymerized. Point mutations or chromosomal deletions in hmgA, which encodes homogentisate 1,2-dioxygenase, result in the accumulation of HGA and subsequent pyomelanin production. My work showed that the pyomelanogenic Pseudomonas aeruginosa clinical isolate PA1111 had a chromosomal deletion of hmgA, while a second pyomelanogenic clinical isolate, DKN343, had a loss of function mutation in HmgA. The 4-hydroxyphenylpyruvate dioxygenase (Hpd) inhibiting compound 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione (NTBC) has been …