Life Sciences Commons^™

Flavodoxin, The Hydrogen Atom Of Flavoproteins: A 19f Nmr Study Of Dynamics And Conformational Changes Utilizing Flavodoxin From Rhodopseudomonas Palustris, Taylor Varner

Theses and Dissertations--Chemistry

Flavodoxin is a small, highly stable protein that contain a single FMN cofactor used to transfer single electrons at low potentials. The organism Rhodopseudomonas palustris contains a long-chain flavodoxin. Long-chain flavodoxins are characterized by a 20 amino acid loop that is proposed to allow interactions with partner proteins. We plan to utilize this protein as a model to build our repertoire with protein fluorination and fluorine NMR in flavoproteins. This tool kit will then be applied to the study of a partner protein of interest that is capable of performing electron bifurcation. We have incorporated m-fluoro tyrosine into flavodoxin and …

Plant-Soil Interactions Dominate Soil Microbial Respiration And Soil Organic Carbon Sequestration In A Subtropical Moist Evergreen Broadleaved Forest In China, Zhijie Yang

Theses and Dissertations--Plant and Soil Sciences

Tropical forest soils contain one-third of global soil carbon (C). The warm and moist climate in tropical forests leads to rapid soil organic carbon (SOC) decomposition, with the highest soil microbial respiration rates in the world, so even a slight change in soil C and microbial respiration could affect atmospheric carbon dioxide concentration. However, there remains a lack of understanding of the mechanisms driving microbial respiration in tropical forests, due to different climate and biophysical drivers compared to temperate or boreal forests. Furthermore, forest conversions (from natural forests to plantations) are most widespread in tropical regions, leading to a loss …

Controlled Release Of Fibroin Repeat-Tagged Proteins From Silk Hydrogel, Jaturong Promsuk

Chulalongkorn University Theses and Dissertations (Chula ETD)

Protein-based biologic drugs such as growth factors are playing important roles in modern medicine. These proteins can be used to stimulate local tissue repair. Growth factors are often incorporated into a scaffold, such as a hydrogel, and then put into the desired area of the body to achieve therapeutic effects. However, diffusion of these growth factors outside of the designated area or inappropriate release time can have deleterious consequences, such as inducing tumors in other areas of the body or uncontrolled inflammation. Therefore, this project aims to create a model to study diffusion of protein from hydrogel by using green …

Protective Effects Of Tiger Milk Mushroom (Lignosus Rhinocerus) Extracts On Oxidative Stress-Induced Neurotoxicity And Aging In Ht22 Cells And C. Elegans, Parinee Kittimongkolsuk

Chulalongkorn University Theses and Dissertations (Chula ETD)

Lignosus rhinocerus (LR) or Tiger Milk Mushroom, a fork medicinal mushroom, has been reported for several pharmacological effects including asthma treatment, anti-inflammatory, anti-proliferative, immuno-modulating effects, promote neurite outgrowth in PC-12 cells, anti-HIV-1 activity, and antioxidants properties. However, the antioxidant properties have only focus on in vitro and no or few studies have reported their protective effects in mouse hippocampal (HT22) cells and Caenorhabditis elegans (C. elegans). This study aims to investigate the neuroprotective effect of three extracts of LR against oxidative stress in both HT22 cells and. C. elegans as well as longevity in C. elegans. In HT22 cells, we …

Antifungal Defense Molecules From Bacterial Symbionts Of North American Trachymyrmex Ants, Georgia Scherer

CMC Senior Theses

Defensive symbioses, in which microbes provide molecular defenses for an animal host, hold great potential as untapped sources of therapeutically useful antibiotics. Fungus-growing ants use antifungal defenses from bacterial symbionts to suppress pathogenic fungi in their nests. Preliminary chemical investigations of symbiotic bacteria from this large family of ants have uncovered novel antifungal molecules with therapeutic potential, such as dentigerumycin and selvamicin.

In this study, the bacterial symbionts of North American Trachymyrmex fungus-growing ants are investigated for antifungal molecules. Plate-based bioassays using ecologically-relevant fungal pathogens confirmed that these bacteria have antifungal activity. In order to purify and identify the antifungal …

The Effects Of Signaling Molecules To Agrobacterium Tumefaciens Tumorigenesis At Plant Wound Sites, Nicole Walker

Honors Theses

Agrobacterium tumefaciens can live independently within soil before infecting its host (McCullen and Binns, 2006). The bacterium infects the plant through the rhizosphere, an area of soil along the plant root surface which contains microorganisms, making up the “external metabolome” (Bais et al. 2006). As these bacteria live in the rhizosphere, they are exposed to a multitude of chemical signals. These chemical signals include several conditions that must be met before the virulence machinery is expressed. These conditions include signaling from phenols and sugars as well as low PO4 levels and low pH. Upon wounding, sugars and phenols are released …

Evaluating A Culprit: A Review Of The Biochemical Mechanisms Of Non-Celiac Gluten Intolerance, Callie Cinque

Honors Theses

Non-Celiac Gluten Intolerance (NCGI) has evaded biological and chemical mechanisms since it was first reported in 1978. Celiac Disease, a multi-system immune mediated disorder has long served as the most similar counterpart to NCGI. Less cases of celiac disease are going undiagnosed reflecting an improvement on sensitive and decisive diagnostic tools. However, NCGI has been significantly growing as a diagnosis over the past decade with little match or similarity to the markers found in celiac disease patients. Critical evaluation of NCGI theories, experimentation, and existing hypotheses is necessary. This review seeks to synthesize multiple disciplines of gluten research and enhance …

Paper-Based Point-Of-Care Tools For Blood Testing, Xuefei Gao

Graduate Theses, Dissertations, and Problem Reports

Early detection of malignant disease is crucial for timely diagnosis and effective medical intervention, which significantly increases survival rates and reduce financial burden on patients. Biomarkers are becoming increasingly important in detection of malignant diseases, because they can be employed for indicating diseases, predicting risks and monitoring the progression of diseases. In addition, biomarkers show up at early stages of diseases in human tissues and fluids (e.g., blood, urine and saliva), which shows great promise for early disease detection. In this dissertation, paper-based lateral flow strips (PLFSs) have been developed for the detection of disease biomarkers, including protein biomarkers and …

Expression, Purification, And Characterization Of Recombinant Human Npas2, Brenda Moreno

Open Access Theses & Dissertations

Many living organisms have biological clocks known as circadian rhythms that control various physiological and behavioral processes, tailored with the day and night. At the cellular level, the circadian clock drives daily rhythms with a transcriptional-translational feedback loop (TTFL) established by multiple transcription factors and genes under their regulation. Circadian Locomotor Output Cycles Kaput (CLOCK) and Brain and Muscle ARNT-like protein 1 (BMAL1) are two of those transcription factors, initiating the TTFL by forming a heterodimeric complex that bind to DNA promoters. Neuronal PAS domain protein 2 (NPAS2) is a functional analog to CLOCK. Less knowledge about NPAS2, in contrast …

Exploring Substrate Specificity Of Fructose Transporters En Route To Glut Specific Probes For Biochemical And Biomedical Applications, Vagarshak Vigenovich Begoyan

Dissertations, Master's Theses and Master's Reports

Carbohydrate Transporters (GLUTs) are responsible for the transportation of sugars into the cell and have been of great interest in research for decades. Alterations or mutations that result in overexpression of GLUTs have been linked to a great number of diseases including, obesity, diabetes, and cancer. Differentiation between transporters has been shown to be incredibly difficult due to the highly conserved nature of the transporter structure, and thus specific targeting of transporters has proven a difficult challenge. Additionally, the GLUTs have been shown to high flexibility in their conformations, so it is difficult to determine what can or cannot pass …

Virus Purification Framework And Enhancement In Aqueous Two-Phase System, Pratik Umesh Joshi

Dissertations, Master's Theses and Master's Reports

Viral infections regularly pose detrimental health risks to humans. Preventing viral infections through global immunization requires the production of large doses of vaccines. The increasing demand for vaccines, especially during pandemics such as COVID-19, has challenged current manufacturing strategy to develop advanced unit operations with high throughput capability. Over the decade, the upstream processing responsible for synthesizing viral products in cell cultures has shown significant success in yielding high titers of viruses and virus-like particles. The progress in the upstream stage has now shifted the bottleneck to the downstream processing (DSP). Overlooked for decades, the DSP responsible for viral product …

Therapeutic Dna: Delivery And As A Delivery Vehicle, Natalie J. Holl

Masters Theses

"A review of gene delivery methods and gene editing methods, as well as original research utilizing DNA as a delivery vehicle is presented in the following thesis. Thousands of diseases have been linked to genes. Gene therapy, either delivering therapeutic genes or editing DNA bases, has arisen as a treatment option with the potential to cure diseases, rather than just ease symptoms. Genes and editing tools need to be delivered to cells for these therapies to be effective and many techniques have been developed to address the issue of delivery. Nonviral and viral methods have been used to deliver nucleic …

Synthesis And Biophysical Analysis Of The Cell-Penetrating Peptide Penetratin And Its Aromatic Analogues, An Le

Theses and Dissertations (Comprehensive)

Cell-penetrating peptides (CPPs) interact with biological membranes, undergo cellular intake/uptake, and may act as potential drug delivery agents. Understanding the molecular interactions of these peptides with membranes contributes to gaining a better knowledge of their potential use in medical and pharmaceutical applications to improve human health. The current research focuses on understanding the mechanisms of a CPP in interaction with different model phospholipid membranes. The peptide penetratin (primary sequence: RQIKIWFQNRRMKWKK) is an example of a CPP that can interact with and pass through biological membranes. The current thesis provides spectroscopic and calorimetric evidence that penetratin associates with lipid membranes. Circular …

Functional Characterization Of Proteins From A Putative Chondroitin Sulfate A Degrading Operon And The Crystal Structure Of An Exo-N-Acetylgalactosamine-4-Sulfatase From Tannerella Forsythia, Peter Nguyen

Theses and Dissertations (Comprehensive)

Advanced periodontitis has been shown to have strong associations with the residence of a bacterial triad of Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola also known as the red complex. These bacteria were previously thought to be asaccharolytic, making them better suited to survive in deep in periodontal pockets where access to carbohydrates from food is limited. Information gathered from genomic analysis suggested that upregulation of an operon (BFO2285-BFO2295, BFO3043) could confer the capacity for T. forsythia to degrade chondroitin-4-sulfate, the principle glycosaminoglycan of alveolar bone tissue. Bioinformatics analysis suggested the operon was similar to an operon found in Bacteroidetes …

Structural And Functional Characterization Of Cazyme And Cazyme-Related Proteins From: Bacteroides Thetaiotaomicron And Porphyromonas Gingivalis: Two Abundant Colonizers Of The Human Microbiome, James Stevenson

Theses and Dissertations (Comprehensive)

The human body consists of approximately 30 trillion cells, while non-human microbes that reside on and within the body outnumber human somatic cells by a factor of 1.3 – 2.3. The interplay between our cells and those of the colonizing microorganisms affect physiology in a multitude of ways, both beneficial and detrimental. Microbes found in the oral cavity, such as the Red Complex member Porphyromonas gingivalis, are associated with pathology, namely periodontal diseases including gum deterioration, tooth decay, and loss of underlying alveolar bone. At the other end of the gastrointestinal tract, microbes such as Bacteroides thetaiotaomicron are found …

Characterization Of The Influences Of Human Cytomegalovirus Glycoprotein O (Go) Expression On Gh/Gl Complexes Assembly And Its Polymorphisms On Cell-Free And Cell-To-Cell Spread, And Antibody Neutralization., Le Z. Day

Graduate Student Theses, Dissertations, & Professional Papers

Human cytomegalovirus (HCMV) is widely spread throughout the world and immunocompromised individuals can suffer severe diseases from HCMV infection. Once the infection is established, HCMV can spread through the body and infect many major somatic cell types. The glycoproteins H and L (gH/gL) on HCMV envelope can be bound by either gO or the UL128-131 proteins to form complexes gH/gL/gO and gH/gL/UL128-131 that are critical for viral entry and spread, and these two complexes are important targets of neutralizing antibodies. Strains of HCMV vary considerably in the levels of gH/gL/gO and gH/gL/UL128-131. gO is one of the most diverse loci …

Seasonal Variation In Mitochondrial Bioenergetics Of The Bluegill Sunfish, Lepomis Macrochirus, From A Shallow Midwest River, Derick Isaac Lamptey

Masters Theses

As average global temperature increase, the frequency and magnitude of extreme temperatures in shallow aquatic ecosystems are more ubiquitous. In order to understand how these changing thermal regimes affect aquatic ectotherms, it is essential to develop studies evaluating the response of ectotherms to seasonal fluctuating thermal regimes. Previous studies on fluctuating temperature regimes have reported an increased physiological stress leading to morphological, behavioral and biochemical adaptations. From the latter, the adaptive capacity and seasonal performance associated with optimal function of the oxidative phosphorylation system (OXPHOS) are key for species persistence. However, studies on this matter are scarce. This study explores …

Investigation Of Glutamate Carboxypeptidase Ii Roles In The Folate Cycle Dependent Reproduction And Development Of Caenorhabditis Elegans, Jessica M. Derham

Masters Theses

Glutamate Carboxypeptidase II (GCPII) is a transmembrane, zinc metallopeptidase that is expressed in a wide range of organisms, including roundworms, mice, and humans. In humans, GCPII is primarily expressed in the prostate, kidneys, small intestine, and central nervous system. Within the small intestine, the expected function of GCPII is to aid in the absorption of dietary folate from the intestinal lumen. GCPII cleaves excess glutamates from folate to yield monoglutamated folate which is then readily transported into the enterocyte. Folate can then be used through the one carbon metabolic cycle for the synthesis of nucleotides, conversion of homocysteine to methionine, …

Application Of Β-Chloroenals: One-Pot Syntheses To Create Highly Variable, Functional, And Biologically Interesting Molecules, Julia Powell Siewert

Honors Theses

Pyrroles and pyrazoles are privileged structures which provide a molecular framework found in many different classes of bioactive compounds, thus rendering their syntheses useful in pharmaceutical drug development. Additionally, being able to selectively create these molecules with interesting substituents allows for different pharmacological and biological activities, such as antitumor and antibiotic effects. Our group has used β-chloroenals in the application of many different unique synthetic strategies in the past, and here I show that chloroenals can be used to synthesize novel 1,2,5 trisubstituted pyrroles as well as 1,5 di- and 1,4,5 trisubstituted pyrazoles in fewer steps and with milder conditions …

Identifying Metabolites As Markers Of Fatigue In Athletes, Christopher D. George

Honors Theses

Fatigue in athletes caused by inadequate rest and other stressors can be severely detrimental to their health, and yet there is no reliable way to measure and track fatigue. Many classical measures of fatigue like body mass and resting heart rate are not reliable ways to track the physical fatigue of an athlete because they can change with many factors. In addition, it is not known how academic stress on top of physical stress affects fatigue. Metabolomics is a relatively new area of study and tracking metabolites offers the possibility to produce biomarkers to assess fatigue. This study collected saliva …

Role Of Ciliary Proteins Adp Ribosylation Factor Like Gtpase 13b (Arl13b) And Bardet-Biedl Syndrome-8 (Bbs8) In Photoreceptor Outer Segment Morphogenesis, Maintenance, And Viability, Tanya L. Dilan

Graduate Theses, Dissertations, and Problem Reports

Photoreceptor neurons are modified primary cilia with an extended ciliary compartment known as the outer segment (OS). The mechanisms behind the elaboration of photoreceptor cilia, OS morphogenesis, and maintenance remain poorly understood. In this work, we focused on dissecting the role of two ciliary proteins, the small GTPase ADP-ribosylation factor-like GTPase 13B (ARL13B) and Bardet-Biedl Syndrome-8 (BBS8) in the context of photoreceptor biology. Both BBS8 and ARL13B are linked to defects in ciliogenesis (cilia development) and Retinitis Pigmentosa (vision loss). ARL13B is implicated in regulating ciliary length, and BBS8 is part of the Bardet-Biedl Syndrome complex (BBSome); the BBSome is …

Evaluation Of Salmonella Enteritidis And Escherichia Coli O157:H7 Volatile Organic Compound Analysis For Food Safety Monitoring: A Preliminary Study, Jessica E. Lemley

Graduate Theses, Dissertations, and Problem Reports

Food borne illness has the potential to impact every point of production from producers to consumers. Food product recalls are common due to the presence of food borne illness causing bacteria. Detection of volatile organic compounds (VOC) have the potential to reduce the number of illnesses and recalls through application of additional monitoring of food products like those most commonly recalled. The objective of this research was to 1) explore the use of closed-loop headspace analysis coupled with gas chromatography-mass spectrometry (GC-MS) on food borne illness causing bacteria and 2) examine the VOC profile of Escherichia coli serotype O157:H7 in …

Proteomic Analysis Of Fetal Rat Neural Stem Cells After Treatment With Hericium Erinaceus, Bright Adam Test

University of the Pacific Theses and Dissertations

The fungus, Hericium erinaceus, has outstanding chemical properties, displaying health benefits in digestive, hepatic, and nervous tissues. Its ease of accessibility and use makes it one of the most common substances used for treatment in Eastern medicine. More and more recent research is confirming the incredible health benefits of this fungus, especially the impact that is seen on nervous tissue growth and recovery post-treatment. Such neurite outgrowth and myelin sheath regeneration could illustrate the beginning of the cure to lifelong neurodegenerative diseases such as Multiple Sclerosis. In this first-of-its-kind study, we cultured and differentiated fetal rat neural stem cells while …

Effect Of Mptp On Α-Synuclein Spreading, Accumulation And Toxicity In Mice With Intrastriatal Innoculation Of Human Α-Synuclein Preformed Fibril, Madiha Mohieldin Merghani

Theses

Parkinson’s disease (PD) is the second most common progressive neurodegenerative disorder, characterized by abnormal accumulation of interneuronal inclusions of α-synuclein (α-syn) known as Lewy bodies (LBs) and Lewy neurites (LNs), and significant loss of dopamine neurons in the substantia nigra pars compacta (SNc) region of the brain. Recent evidence suggests that intrastriatal inoculation of α-syn Preformed Fibril (PFF) into mice induces PD-like LBs and LNs pathology formed by aggregated α-syn in anatomically interconnected brain regions which needs several months of incubation to develop the pathology. In the present study, we have evaluated the effect of different doses of 1-methyl, 4-phenyl, …

Deconvolution Tools For Extracting Insight From Challenging Two-Flavin Systems, Dallas Michael Bell

Theses and Dissertations--Chemistry

Flavoproteins have long been explored for their ubiquity among a number of metabolic and energetic reactions. The flavin cofactor has the inherent benefit of distinct spectral changes associated with redox transitions; however, the double-edged sword is incurred as these distinct signatures overlap and take up much of the UV-vis spectral range. Therefore, it is crucial to create a method to demarcate the expressed redox transitions for studying these systems. The first portion of these studies discusses the creation of a program that deduces spectra for redox transitions in a single-flavin containing model protein: flavodoxin. The latter portions discuss the application …

Characterization Of The Dyrk1a Protein-Protein Interaction Network, Varsha Ananthapadmanabhan

Theses and Dissertations

Human Dual specificity tyrosine (Y)-Regulated Kinase 1A (DYRK1A) is a protein kinase encoded by a dosage-dependent gene. An extra copy of DYRK1A contributes to Down syndrome (DS) pathogenesis while loss of one allele causes severe mental retardation and autism. DYRK1A is involved in phosphorylation of several proteins that regulate cell cycle control and tumor suppression. However, the function and regulation of this kinase is not well understood and current knowledge does not fully explain dosage-dependent function of this important kinase. Our previous proteomic studies identified several novel DYRK1A interacting proteins including RNF169, FAM117B, TROAP, LZTS1, LZTS2 and DCAF7. In this …

Characterization Of The Tsc/Dyrk1a Interaction, Supriya Joshi

Theses and Dissertations

The Tuberous sclerosis complex (TSC) includes TSC1, TSC2 and the TBC1D7 subunits that together function as a principal inhibitor of the mTOR protein kinase complex 1 (mTORC1). mTORC1 is a master regulator of cell growth and proliferation that responds to signaling cues such as growth factors and nutrient availability. Proteomic studies in our lab revealed an interaction between the TSC subunits and DYRK1A, a ubiquitous protein kinase encoded by a gene located in the Down syndrome (DS) region on human chr21. In this study, we sought to validate the interaction of the TSC components with DYRK1A and to determine the …

Novel Post-Translational Modification And Function Of Fus: The Relevance To Amyotrophic Lateral Sclerosis, Alexandra Arenas

Theses and Dissertations--Toxicology and Cancer Biology

Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease characterized by the preferential death of motor neurons. Approximately 10% of ALS cases are familial and 90% are sporadic. Fused in Sarcoma (FUS) is a ubiquitously expressed RNA binding protein implicated in familial ALS and frontotemporal dementia (FTD). FUS is ubiquitously expressed in cells and has a variety of functions in the nucleus and cytoplasm. FUS mutations in the nuclear localization sequence (NLS) causes mislocalization of FUS in the cytoplasm, where it can undergo liquid-liquid phase separation and become stress granules or protein inclusions. Although FUS inclusion bodies can be found in …