Biochemistry, Biophysics, and Structural Biology Commons^™

Deciphering The Role Of Glycine134 In The Human Acidic Growth Factor-1’S Binding To Heparin, Adam W. Burroughs

Chemistry & Biochemistry Undergraduate Honors Theses

Human acidic fibroblast growth factor 1 (FGF-1) is a potent modulator of cell survival and exhibits a universal role in various physiological processes. Though potent, FGF-1 unbound to heparin is known to show a poor thermal stability and a relatively short in vivo half-life. Much is known about the structure and relation of FGF-1 with heparin yet there is still unknown information regarding the exact role of heparin in stabilizing FGF-1. Thus, the aim of this study is to mutate glycine at position 134 to glutamic acid in wild type FGF1. G134 is located in the heparin binding pocket, thus …

The Effects Of Pulmonary Hypertension In Diabetic Zucker Rats On Pulmonary Vascular Contraction And Right Ventricular Size, Michael Franzetti

Biological Sciences Undergraduate Honors Theses

Pulmonary hypertension is a life-threatening disease that is identified by a resting mean pulmonary arterial pressure over 25 mmHg and established by right heart catheterization (Saglam et al., 2015). The symptoms commonly associated with pulmonary hypertension include fatigue, dyspnea, syncope, and chest pain, which severely limit quality of life in diagnosed patients (Saglam et al., 2015). According to recent studies in patients with pulmonary hypertension, the one, three and five year mortality rates are 8%, 25%, and 34%, respectively (Burudpakdee, Shah, Joish, Divers, & Yaldo, 2014). Although some advances have been made in therapies for pulmonary hypertension, the prognosis is …

The Ccaat-Binding Factor Dependent Regulation Of The Oxidative Stress Response In Candida Albicans, Ananya Chakravarti

Graduate Theses and Dissertations

The success of Candida albicans as an opportunistic human pathogen has been attributed to several factors, including the ability to survive in limiting iron environments and the ability to evade the respiratory burst of human macrophages and neutrophils. The goal of this research is to elucidate the role of the CCAAT-binding factor in the oxidative stress response of Candida albicans. Prior whole genome microarray studies performed in our lab compared the gene expression of a wild type Candida albicans strain versus a hap5Δ strain under iron-limiting growth conditions. Among the differentially regulated genes, CTA1, encoding catalase, had a four-fold higher …

Synthesis Of Microgel Polymers As Catalysts, Hannah N. Miller

Chemistry & Biochemistry Undergraduate Honors Theses

New developments in organic synthesis show promise in achieving the best catalytic properties for the hydrolysis of glycosidic bonds through microgel polymers and transition metal complexes. A monomer mix of ethylene glycol dimethacrylate, butyl acrylate, and styrene form miniemulsion polymers after sonication and exposure to UV light. Gravimetrical analysis is used to determine the most suitable polymerization conditions by performing experiments at varying pH values, temperatures, monomer amounts, initiator amounts, and lamp heights. The final data show that the best polymerization conditions are a pH of 10.50 at 0°C with a high monomer ratio, 20% initiator amount, and a lamp …

Gene Expression And Physiological Analysis To Study Differences Between Oryza Sativa Cultivars Susceptible And Resistant To Chalky Grain Formation Subjected To High Nighttime Temperatures., Nicholas Lawson

Graduate Theses and Dissertations

Starch composition and grain quality of rice is greatly influenced by genotype and environmental factors. The detrimental effects of high nighttime temperatures on rice yield and quality has recently become apparent, with some of the warmest average nighttime temperatures being recorded in the past few years. One of the most notable effects of this stress, an increase in grain chalk formation, correlates with a decrease in quality. This effect varies greatly between cultivars as some show less temperature-sensitive quality reduction than others. The goal of this research is to elucidate fundamental changes that occur in developing plants and grains as …

Genotoxicity Of Graphene In Escherichia Coli, Ananya Sharma

Graduate Theses and Dissertations

Rapid advances in nanotechnology necessitate assessment of the safety of nanomaterials in the resulting products and applications. One key nanomaterial attracting much interest in many areas of science and technology is graphene. Graphene is a one atom thick carbon allotrope arranged in a two-dimensional honeycomb lattice. In addition to being extremely thin, graphene has several extraordinary physical properties such as its exceptional mechanical strength, thermal stability, and high electrical conductivity. Graphene itself is relatively chemically inert and therefore pristine graphene must undergo a process called functionalization, which is combination of chemical and physical treatments that change the properties of graphene, …

Oxidative Stress Response In Archaea: Elucidation Of Oxidant Sensing And Tolerance Mechanisms In Methanosarcina Acetivorans, Matthew Edward Jennings

Graduate Theses and Dissertations

Methanogens are archaea possessing a conserved metabolic pathway which produces methane. Many of the enzymes in the methanogenesis pathway are Fe-S proteins, meaning methanogens are sensitive to conditions which disrupt Fe-S clusters. Molecular oxygen is capable of disrupting Fe-S clusters through oxidation of the iron atoms. Furthermore, reduced iron can facilitate the production of reactive oxygen species (ROS), meaning methanogens must possess antioxidant mechanisms. Detection and eradication of ROS is important for all cells, due to the potentially fatal consequences of unchecked oxidation. This dissertation presents two separate projects investigating mechanisms the model methanogen Methanosarcina acetivorans possess for dealing with …

Proteomic Characterization Of Eggshell Membranes And Their Effect On Poultry Physiology And Immunity, Sarbjeet Kaur Makkar

Graduate Theses and Dissertations

The use of antibiotics in poultry growth and disease control has led to antibiotics resistant problem in human beings, which is a big concern among consumers. With the necessity for judicious use of antibiotics in poultry production, alternative strategies to improve disease resistance in poultry production are necessary. The research is more inclined towards using the natural products available to grow healthier and antibiotic free meat animals. In the context of exploring natural and sustainable resource of alternative to antibiotics, the biochemical milieu of eggshell membranes (ESM) were analyzed by using mass spectrometry techniques including matrix assisted laser desorption ionization …

Investigation Of The Inherent Chemical, Structural, And Mechanical Attributes Of Bio-Engineered Composites Found In Nature: Alligator Gar’S Exoskeleton Fish Scales, Wayne Derald Hodo

Graduate Theses and Dissertations

The U.S. Army has determined a huge cost savings of up to 51% can be accomplished by reducing the gross vehicle weight, for their personnel carrier, by 33%. To cut cost, composite materials are needed. Man-made composites can have superior material properties (high-strength, high-fracture toughness, and lightweight), but they are prone to delamination at the glued-layered interface. In contrast, fish scale is a natural composite that has the same material properties and, additionally, tend not to delaminate.

The focus of this study was to learn how nature integrates hard and soft materials at each length scale to form a layered …

Blackberry Virosome: A Micro And Macro Approach, Archana Khadgi

Graduate Theses and Dissertations

Viruses pose a major concern for blackberry production around the world with more than 40 species known to infect the crop. Virus complexes have been identified recently as the major cause of plant decline with blackberry yellow vein disease (BYVD) being the most important disease of the crop in the Southern United States. The objective of this research was to study the blackberry virosome in both the macro and micro scale. The large scale approach involves identification of the major viruses known to be associated with BYVD in the Southern United States as well as the identification of other viruses …

Leptin Regulates The Expression Of Autophagy-Related Genes In Chickens, Peter Olawale Ishola

Graduate Theses and Dissertations

Autophagy or cellular self-digestion, a lysosomal degradation pathway that is conserved from yeast to human, plays a key role in recycling cellular constituents, including damaged organelles. It also plays a pivotal role in the adaptation of cells to a plethora of distinct stressors including starvation. Autophagy has been extensively studied in mammals and yeast, but little is known in avian species. Thus, the major objective of the present study was to determine the effects of leptin on autophagy-related genes in chicken hypothalamus, muscle and liver. Leptin is an adipocytokine that is mostly produced by white adipose cells in mammals (as …

Optimizing A Luciferase-Based Tool For Studying The Effects Of Fatty Acid Desaturase 7 On Singlet Oxygen Accumulation In Arabidopsis Thaliana, Abeer Muhammedali Alnasrawi

Graduate Theses and Dissertations

In plants, reactive oxygen species (ROS) are generated as a byproduct of normal metabolism, as well as in response to adverse conditions such as light stress, extreme temperatures, and exposure to pests and pathogens. Singlet oxygen (1O2) is a ROS that is formed during photosynthesis in photosystem II (PSII) of the chloroplasts. Levels of 1O2 and other ROS are tightly controlled in healthy plants, but some studies suggest that levels of fatty acid desaturase (FAD) activity in Arabidopsis thaliana can influence constitutive and/or stress-responsive ROS accumulation. In this study, a luciferase-based reporter gene that is selectively stimulated by 1O2 (AAA-ATPase: …

Analysis Of Ac/Ds Activation Tagged Mutants In Tomato (Solanum Lycopersicum), Ipeleng Randome

Graduate Theses and Dissertations

Tomato (Solanum lycopersicum) is a crop of immense economic and nutritional importance worldwide and also a good model organism for genomic studies of other dicot species. The recent completion of the tomato genome sequence is a great milestone towards learning about the tomato genome. Elucidation of the function of the different genes using different functional genomic tools is therefore important in adding to this resource. To this end, we have developed an Ac-Ds transposon ‘activation tagging’ (ATag) system to be able to transpose transposon inserts, bearing a strong 35S-enhancer element, all around the genome. An Ac-Ds ATag construct was used …

Transcriptomics Of Chicken Primordial Germ Cells, Nhung Thi Nguyen

Graduate Theses and Dissertations

Chicken primordial germ cells (PGCs) are derived from extraembryonic tissue of the embryo and first appear at stage X of development. They enter the bloodstream and migrate to the genital ridge, unite with somatic tissue to form a developing gonad, and then differentiate to sperm or ova (Fujimoto et al., 1976). Understanding molecular features of both male and female PGCs not only clarify the differentiation mechanism of such cells toward different germ lines, but will also help in selecting for highly productive types of commercial chicken. Most previous studies focused on the location of PGCs (Eyal-Giladi et al., 1981; Swift …

Autophagy And Its Potential Role In Stress And Feed Efficiency Using Avian Lines, Alissa Laura Piekarski

Graduate Theses and Dissertations

Autophagy is a highly conserved cellular mechanism that is responsible for the degradation and recycling of damaged organelles. Recently, autophagy has been involved in critical roles during overall development of the organism and degradation of damaged cellular components. This pathway has witnessed dramatic growth in the last few years and has been extensively studied in yeast and mammals, however, there is a paucity of information in avian (non-mammalian) species. First, we characterized genes involved in the autophagy pathway in male and female Jungle Fowl to determine gender and tissue specific differences. Secondly, tissue and genotype differences in Japanese quail selected …

Response Of Bacterial Cells To Fluctuating Environment, Sudip Nepal

Graduate Theses and Dissertations

We have studied morphological and genomic variations occurring in a mesophilic bacterium Escherichia coli (E. coli) in a wide range of continuous and fluctuating hydrostatic pressures. For all the studies here the temperature is maintained at 37◦C, the optimal growth tem- perature of E. coli at atmospheric pressure. Cell division is inhibited at high hydrostatic pressures resulting in an increase of cell length. The increase of cell-length depends on the extent and duration of the stress applied on bacterial cells. We have studied the effect of high pressure stress in three different conditions – (i) Wild-type cells (almost no genetic …

In Vivo Periodical Monitoring Of Immune Cell Infiltration In Response To Feathers And Intramuscular Injection Of Ionps Using The Pulp (Dermis) Of Growing Feathers As Test Site Tissue In Chickens, Olfat Taleb Alaamri

Graduate Theses and Dissertations

The applications of nanoparticles are growing, but little is known about their interactions with the immune system as most studies did not use suitable in vivo test systems. The purpose of these studies was to investigate the utility of chicken growing feathers as an in vivo test site for iron oxide nanoparticle (IONP) interactions with the immune system. The first objective of this study was to monitor leukocyte infiltration into the growing feather pulp upon the administration of IONP and IO-mIgG preparations. The second objective was to test the utility of IONPs as vaccine adjuvants by monitoring primary and memory …

Understanding The Causal Agent Of Rose Rosette Disease, Patrick Louis Di Bello

Graduate Theses and Dissertations

A number viruses are known to infect roses, ranging from those in the genera Nepovirus, and Ilarvirus, which have been reported since the inception of rose virology, to recently discovered viruses in the genera Carmovirus, Closterovirus, Emaravirus, Luteovirus, Rosadnavirus, and Potyvirus. Of the viral diseases in rose, arguably the most damaging is Rose rosette (RRD), which is associated with the Emaravirus, Rose rosette virus (RRV). The objective of this thesis is to fill in the gaps in knowledge on the epidemiological aspects of RRD and RRV. There has been significant progress in the epidemiology of the RRD agent prior to …

Using Peptoids To Build Robust, Efficient Microarray Systems, Dhaval Sunil Shah

Graduate Theses and Dissertations

Recent studies have shown microarrays to be indispensable for various biological applications, allowing for high-throughput processing and screening of biological samples such as RNA, DNA, proteins and peptides using a small sample volume (< 1 µL). Peptoids (poly-N-substituted glycine oligomers) can be used as a substitute for antibodies as capture molecules, as well as coatings for slides in antibody microarrays. The ease of synthesis of peptoids, high customizability with desired bioactivity, and speed of synthesis allows us to build a diagnostic system with a large dynamic range that can detect biomolecules from a minimal sample size. In this study, peptoid-based antibody mimics are designed to have both structural and functional features similar to those of antibodies, including a stable constant region (scaffolding) and a variable region for protein recognition. Peptoids previously screened via combinatorial library synthesis to be specific to bind Mdm-2 (mouse double minute 2 homolog) and GST (gluthathione S-transferase), have been synthesized. The protein recognition peptoids have been conjugated to PEG (polyethylene glycol) molecules with modified end groups; an amine group on one end that allows for immobilization and orientation on the slide, and an azide group on the other end that will allow for attachment to the peptoid through “click chemistry”. The number of capture molecules printed on the slides can be increased by making the available surface area of the slide larger via coating with microspheres. We have determined that partially water soluble peptoids that are also helical, can self-assemble into microspheres. Sequences have been developed that can consistently produce uniform microsphere coatings on slides that increase the overall surface area. A high surface area corresponds to a higher number of binding sites, and therefore a more sensitive system. The work done has shown that slides may be successfully coated in order to potentially improve the detection system.

Characterization Of The Effects Of Bfgf And Gsk3Β Inhibitors On Embryoid Bodies Derived From Human Pluripotent Stem Cells, Jonathan Earls

Graduate Theses and Dissertations

Embryoid body (EB) formation is a common first step in many human pluripotent stem cell (hPSC) differentiation protocols. Previous work suggests that EBs are sensitive to growth factor withdrawal if they are derived from hPSCs maintained in feeder independent media such as mTeSR1. To promote cell survival, EBs generated from mTeSR1-adapted hPSCs are sometimes cultured in a medium that contains basic fibroblast growth factor (bFGF), a trophic factor often used in hPSC cultures to maintain self-renewal. This distinguishes feeder independent hPSCs from feeder dependent hPSCs. The purpose of this study was to characterize the effects of bFGF as well as …

Designing Fret Assays To Study Electrostatic Interactions Pertaining To The Binding Of Intrinsically Disordered Proteins, Ashley Ann Howard

Graduate Theses and Dissertations

Fibroblast growth factor receptor plays a major role in several biological processes. Without FGFR, a human cannot live. FGFR is involved in cell differentiation and wound healing. Of course, if FGFR signaling becomes unregulated, it causes severe distress in the body. Several cancers are contributed to high signaling levels, as well as developmental conditions like rickets and Kallmann’s syndrome. FGFR is thought to undergo an auto-inhibition (or self-regulatory) process in order to try to facilitate regulation. The exact method of this inhibition is currently unknown, but is proposed to involve the unstructured acid box region of FGFR. We developed a …

Comparison Of Different Modulators That Affect Macrophage Activation In Vitro, Alda Alexa Díaz Pérez

Graduate Theses and Dissertations

Inflammation is known as a mechanism to regulate and control infections as well as promote tissue repair. Macrophages (Mф) are known to be a major cell type in the initiation, sustainability and resolution of inflammation. Moreover, Mф are essential for the remodeling process that is also known as the wound healing response. The objective of this research was to compare five modulators (acetylsalicylic acid (ASA), dexamethasone (DEX), prostaglandin E2 (PGE2), iloprost, and resolvin D1 (RvD1) for their anti-inflammatory effects on macrophages in vitro. Then, Mф phenotype in terms of gene expression and secreted cytokine response was determined. Our study compared …

A Morpho-Physiological Analysis Of Diverse Drought Resistant And Sensitive Rice Genotypes To Identify Distinguishing Anatomical Root Phenes, Clinton Philip Greub

Graduate Theses and Dissertations

A set of 37 genotypes selected from a survey of over 200 diverse germplasms, for their diverse drought tolerance mechanisms, were analyzed in this study for root physiological traits that are indicative of different drought resistance mechanisms. The genotypes include controls such as the drought sensitive IR64 and Nipponbare, along with the drought resistant Nagina 22 (N22) and Bengal landraces to identify distinguishing phenotypic features of resistant accessions. Anatomical traits studied include total areas of the cross-sections of the stele, cortex, aerenchyma, cortical-cell, and xylem tissue; the number of aerenchyma, xylem vessels, cortical cell file number and size that together …

Production And Characterization Of Islet Amyloid Polypeptide Using Recombinant Technology, Emilio Duverna

Graduate Theses and Dissertations

Islet amyloid polypeptide (IAPP) also called amylin is an amyloid-forming protein; IAPP is a proteinaceous hormone that comprises 37 amino acid residues. It is secreted along with insulin from the pancreatic β-cells to help it regulate the uptake and removal of glucose in the bloodstream. IAPP has been observed in the amyloid deposits found in pancreatic β-cells of most patients suffering from type II diabetes mellitus. This research project aims at producing recombinant amylin peptide. To achieve this goal, we used the pBAD plasmid vector which we introduced into Escherichia coli to express the peptide. Although the vector was successfully …

Biofilm-Related Materials As Total N-Nitrosamine (Tono) Precursors And Hydroxylamine-Based Interferences In Tono And N-Nitrosodimethylamine (Ndma) Measurements, David Allen Meints

Graduate Theses and Dissertations

In this study, a chemiluminescence-based total N-nitrosamine (TONO) assay was adapted to include a solid-phase extraction (SPE) step to assess the role of biologically derived materials as N-nitrosamine precursors. Methanol was determined to be a suitable solvent for the SPE-TONO assay, with N-nitrosodimethylamine (NDMA) losses estimated to be 30% by SPE alone and an additional 15% from concentration of the methanol extracts by N2-gas blowdown. Three biofilm-derived materials - poly-N-acetylglucosamine (PNAG), Pseudomonas aeruginosa, and tryptophan - were individually chloraminated and dose-response relationships were observed with the SPE-TONO assay, indicating biofilm are potential N-nitrosamine precursors. The role of hydroxylamine - a …

Design Of Antimicrobial Peptide Based On The Heparin Binding Segment Of Fgf-1, Alice Margaret Power

Chemistry & Biochemistry Undergraduate Honors Theses

Fibroblast Growth Factors (FGFs) are heparin-binding proteins known for their involvement in various biological processes such as cell differentiation and wound healing.1 The heparin binding site of FGF-1 displays a unique stretch of positive amino acid sequence and facilitates a very strong binding to negatively charged heparin due to the formation of electrostatic interaction. The aim of this study is to analyze and modify the novel antimicrobial peptide sequence (GST-HB) designed based on the heparin-binding region of FGF-1 as well as other polycationic microbial sequences. These aspects will be examined using various experimental techniques including overexpression of GST-HB, purification using …

Additional Sex Combs-Like 2 (Asxl2): A Psychological Stress-Related Gene In A Chicken Stress Model, Emily Sandra Jacobson

Chemistry & Biochemistry Undergraduate Honors Theses

Chronic stress is a problem not only in terms of its psychological effects but also due to the harmful physiological changes induced in the individual. Although chronic stress is recognized as detrimental and rampant throughout society, the stress system itself remains poorly understood in terms of comprehending which genes are involved in regulating and carrying out stress-induced signals. In a previous study, 57 candidate genes in the stress pathway had been identified based on the presence of stress-associated single nucleotide polymorphisms (SNPs) and differential expression from microarray data. The objective of this study was to investigate Additional Sex Combs-Like 2 …

Devlopment Toward Fluorescence Imaging Of Microdialysis Sampling Diffusion Profiles, Nicholas Edward Baioni

Chemistry & Biochemistry Undergraduate Honors Theses

Microdialysis is a diffusion-based method used to collect analytes from specific locations in the body. Matrix Metalloproteinases (MMPs) are important enzymes located throughout the body that play a role in the breakdown of extracellular matrix. Microdialysis sampling in conjunction with appropriate analysis methods can be used to observe the reactions of MMPs in real time. This project is aimed at determining the activity of porcine pancreatic enzyme, a known substitute for MMP-12, using nanodrop spectroscopy. This is a necessary pre-requisite toward using microdialysis in an in vivo imaging setup. These experiments were performed using Succinyl(Ala)_3-p-nitroanilide (suc(Ala)3-p-NA), 4-nitroaniline, and porcine pancreatic …

Molecular Mechanisms Of Tandem Cbd Of Clostridium Histolyticum, Dawn Weir

Chemistry & Biochemistry Undergraduate Honors Theses

In order to spread infections, bacterial collagenases methodically unravel collagen fibril in tissues. Collagen is the most abundant protein in the body, and can be found in the skin, bone and cartilage [1]. Two collagenases, ColG and ColH, synergistically dismantle collagen fibrils by seeking different weak links in the collagen structure. The collagen-binding domain (CBD) of these collagenases binds to most vulnerable regions in collagen [8]. Without CBDs, collagen fibril cannot be degraded. Cells express collagen receptors in order to anchor themselves, which is a critical step in cell proliferation. Binding sites for some collagen receptors, such as integrin and …

Efforts In Increasing Microdialysis Recovery Rates By Utilizing Bidirectional Flow Capabilites, Jacob Taylor Needham

Chemistry & Biochemistry Undergraduate Honors Theses

Microdialysis sampling involves the collection of biological fluids from tissues or fluid-filled cavities in vivo via diffusion through a semipermeable membrane probe. In order to increase the recovery of fluids for analysis from this process, a new regime was attempted that would allow perfusion fluids to make multiple passes through the probe in order to collect more analyte with each additional pass. This was dubbed the Bidirectional Flow Technique. Dextran-70 solution was used as the perfusion fluid while 100μM Methyl Orange solution was used as the analyte. The experiments were performed in vitro using a fully automated microdialysis ePump capable …