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Phosphotransacetylase And Xylulose 5-Phosphate/Fructose 6-Phosphate Phosphoketolase: Two Eukaryotic Partners Of Acetate Kinase, Tonya Taylor 2015 Clemson University

Phosphotransacetylase And Xylulose 5-Phosphate/Fructose 6-Phosphate Phosphoketolase: Two Eukaryotic Partners Of Acetate Kinase, Tonya Taylor

All Dissertations

Although acetate is a predominant metabolite produced by many eukaryotic microbes, far less attention has been given to acetate metabolism in eukaryotes than in bacteria and archaea. Acetate kinase (Ack), which catalyzes the reversible phosphorylation of acetate from ATP, is a key enzyme in bacterial acetate metabolism. Ack primarily partners with phosphotransacetylase (Pta), which catalyzes the generation of acetyl phosphate from acetyl-CoA, but can also partner with xylulose 5-phosphate/fructose 6-phosphate phosphoketolase (Xfp), which produces acetyl phosphate from either xylulose 5-phosphate or fructose 6-phosphate. The Ack-Pta pathway, found primarily in bacteria, is also present in lower eukaryotes such as the green …


Distinguishing Macrophage Activation States By Mass Spectrometry, Matthias Manfred Knust 2015 University of Arkansas, Fayetteville

Distinguishing Macrophage Activation States By Mass Spectrometry, Matthias Manfred Knust

Graduate Theses and Dissertations

Macrophages are versatile and highly adaptive cells that are involved in a wide range of physiological processes including host defense, homeostasis or regeneration, as well as pathogenesis. They react to their microenvironment, assuming various roles based on chemical and/or physical cues, and can reversibly shift between these so-called activation states. Concurrently, the technique of immunohistochemistry is used to gain spatial information on activated macrophages on tissue sections. The aim of this work was to find mass spectral biomarkers that allow the differentiation of activation states, and establish conditions that can be used in imaging mass spectrometry (IMS) experiments to investigate …


Local Modulation And Measurement Of Macrophage-Derived Bioactive Proteins From Implanted Biomaterials In Rat, Geetika Bajpai 2015 University of Arkansas, Fayetteville

Local Modulation And Measurement Of Macrophage-Derived Bioactive Proteins From Implanted Biomaterials In Rat, Geetika Bajpai

Graduate Theses and Dissertations

Fibrosis around the implanted medical devices is a severe problem that can plague long-term device reliability. Activation of macrophage phenotype (macrophage polarization) has emerged as a new and possible means for reducing fibrosis in the fields of biomaterials and regenerative medicine. Macrophages are phagocytic cells that respond to microenvironmental cues that direct their phenotype. Macrophage activation has been widely studied in mouse and human in the context of tumor biology, yet little information is available regarding how macrophage activation could be used in a biomaterials context. Further, rats rather than mice are the common subjects in biomaterials experiments. A significant …


Beryllium Is A Potent And Unique Gsk-3Β Inhibitor With Potential To Differentially Regulate Glycogen Synthase And Β-Catenin, Ata Ur Rahman Mohammed Abdul 2015 University of Nevada, Las Vegas

Beryllium Is A Potent And Unique Gsk-3Β Inhibitor With Potential To Differentially Regulate Glycogen Synthase And Β-Catenin, Ata Ur Rahman Mohammed Abdul

UNLV Theses, Dissertations, Professional Papers, and Capstones

Glycogen Synthase Kinase-3β (GSK-3β) is an important serine/threonine kinase that phosphorylates/regulates diverse and metabolically important proteins. Some of the important substrates of GSK-3β are glycogen synthase, tau, β-catenin, cyclin D1, axin, c-jun, c-myc, Heat Shock Factor–1, BCL-3, CREB, Histone H1.5, mdm2, p21 (CIP1), pyruvate dehydrogenase and many more. De-regulation of GSK-3β has been implicated in diseases like cancer, Alzheimer’s disease, bipolar disorder and type 2 diabetes mellitus. Currently, GSK-3β is one of the most widely studied proteins and there is a great interest in developing potent and efficient GSK-3β-inhibitors for research as well as therapeutic purposes. We demonstrate that beryllium …


Dlipin-A Link Between Lipid Metabolism, Glucose Homeostasis And Growth In Drosophila Melanogaster, Sandra Schmitt 2015 University of Arkansas, Fayetteville

Dlipin-A Link Between Lipid Metabolism, Glucose Homeostasis And Growth In Drosophila Melanogaster, Sandra Schmitt

Graduate Theses and Dissertations

Lipins are a family of highly conserved proteins found from yeasts to humans. Lipins have dual functions, serving as phosphatidate phosphatase enzymes (PAP) in the synthesis of neutral fats (triacylglycerols, TAG) and as transcriptional co-regulators that affect the expression of genes involved in lipid and fatty acid metabolism. Thus, they play central roles in metabolic control. Disruption of Lipin function has been implicated in lipodystrophy, obesity and insulin resistance. Using dLipin, the Drosophila homolog of Lipin, as a model, I aimed to elucidate the relationship between the two biochemical functions of Lipin and metabolic homeostasis. I discovered there is a …


Role Of C-Kit In The Growth, Migration And Differentiation Of Human Cardiac Progenitor Cells., Bathri Narayan Vajravelu 2015 University of Louisville

Role Of C-Kit In The Growth, Migration And Differentiation Of Human Cardiac Progenitor Cells., Bathri Narayan Vajravelu

Electronic Theses and Dissertations

The belief that adult mammalian heart lacks regenerative potential was challenged by the identification of c-kit positive cardiac progenitor cells (CPCs) in the heart. A recent phase I clinical trial (SCIPIO), has shown that autologous c-kit positive CPCs improve cardiac function and quality of life when transplanted into ischemic heart disease patients. c-Kit is a type III receptor tyrosine kinase and a common stem cell antigen. Stem cell factor (SCF) is the only known ligand for c-kit. Although c-kit is extensively used as an invariable marker of resident CPCs and shown to be important in the context of different cell …


Influence Of A Potentially Destabilizing Central Tryptophan On Transmembrane Helix Domains, Vasupradha Suresh Kumar 2015 University of Arkansas, Fayetteville

Influence Of A Potentially Destabilizing Central Tryptophan On Transmembrane Helix Domains, Vasupradha Suresh Kumar

Chemistry & Biochemistry Undergraduate Honors Theses

The vital role that membrane proteins play in cell mechanisms and the inner workings of disease in our bodies has been universally accepted. Thus, the study of protein-membrane and transmembrane interactions has become increasingly significant in understanding these membrane proteins. Synthetic model peptides, such as GWALP23 (acetyl-GGALW5LALALALALALALW19LAGA-amide)—designed at the University of Arkansas and composed of the amino acid residues of glycine (G), alanine (A), leucine (L) and tryptophan (W) —provide a favorable “host” framework for investigations of the influence of chosen “guest” amino acids. For example, it is of interest to know the consequences of having a third, centrally located, …


Percent Recovery Of Various Analytes Using A Wick Method, Patrick A. Naeger 2015 University of Arkansas, Fayetteville

Percent Recovery Of Various Analytes Using A Wick Method, Patrick A. Naeger

Chemistry & Biochemistry Undergraduate Honors Theses

Finding alternate methods to measure and isolate interstitial fluid in vivo has become a consequence of the limited recovery of analytes when using a probe.1 These probes consist of an inlet constantly bringing perfusate into a semi-permeable membrane where the perfusate is replace with dialysate containing numerous analytes. The dialysate leaves the semi-permeable membrane via an outlet. A wick technique has been shown to be useful in analytical chemistry because it can be used to effectively isolate interstitial fluid and accurately measure the concentrations of various analytes. For long-term applications, wicks can be used to measure the concentrations of cytokines …


Molecular Mechanisms Of Tandem Cbd Of Clostridium Histolyticum, Dawn Weir 2015 University of Arkansas, Fayetteville

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 2015 University of Arkansas, Fayetteville

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 …


Whole Genome Comparison Of A Large Collection Of Mycobacteriophages Reveals A Continuum Of Phage Genetic Diversity, Welkin H. Pope, Charles A. Bowman, Daniel A. Russell, Deborah Jacobs-Sera, David J. Asai, Steven G. Cresawn, William R. Jacobs Jr, Roger W. Hendrix, Jeffrey G. Lawrence, Graham F. Hatfull, Dr. Kathleen Cornely 2015 Providence College

Whole Genome Comparison Of A Large Collection Of Mycobacteriophages Reveals A Continuum Of Phage Genetic Diversity, Welkin H. Pope, Charles A. Bowman, Daniel A. Russell, Deborah Jacobs-Sera, David J. Asai, Steven G. Cresawn, William R. Jacobs Jr, Roger W. Hendrix, Jeffrey G. Lawrence, Graham F. Hatfull, Dr. Kathleen Cornely

Chemistry & Biochemistry Faculty Publications

The bacteriophage population is large, dynamic, ancient, and genetically diverse. Limited genomic information shows that phage genomes are mosaic, and the genetic architecture of phage populations remains ill-defined. To understand the population structure of phages infecting a single host strain, we isolated, sequenced, and compared 627 phages of Mycobacterium smegmatis. Their genetic diversity is considerable, and there are 28 distinct genomic types (clusters) with related nucleotide sequences. However, amino acid sequence comparisons show pervasive genomic mosaicism, and quantification of inter-cluster and intra-cluster relatedness reveals a continuum of genetic diversity, albeit with uneven representation of different phages. Furthermore, rarefaction analysis shows …


Repsa-Directed Identification Of Dna-Binding Specificity For Orphan Transcription Factors, Kamir Hiam 2015 Kennesaw State University

Repsa-Directed Identification Of Dna-Binding Specificity For Orphan Transcription Factors, Kamir Hiam

Symposium of Student Scholars

The function of many genes and the biological roles of their encoded products are still not well characterized. Given the sequence-specific DNA-binding properties of transcription factor proteins, it is possible to purify them, identify the responsible polypeptide(s), determine their consensus binding sequences, and identify their genomic binding sites. Thus, one can go from cellular extract to proposed biological regulatory roles in relatively short order. Our goal is to identify and characterize orphan DNA-binding proteins in the model organism E. coli K12 using the novel combinatorial technique, REPSA (Restriction Endonuclease Protection Selection Amplification), as well as further develop the REPSA to …


Nmr Spectroscopy Reveals The Presence And Association Of Lipids And Keratin In Adhesive Gecko Setae, Dharamdeep Jain, Alyssa Y. Stark, Peter H. Niewiarowski, Toshikazu Miyoshi, Ali Dhinojwala 2015 University of Akron Main Campus

Nmr Spectroscopy Reveals The Presence And Association Of Lipids And Keratin In Adhesive Gecko Setae, Dharamdeep Jain, Alyssa Y. Stark, Peter H. Niewiarowski, Toshikazu Miyoshi, Ali Dhinojwala

Polymer Science Faculty Research

Lipid and protein aggregates are one of the fundamental materials of biological systems. Examples include cell membranes, insect cuticle, vertebrate epidermis, feathers, hair and adhesive structures known as ‘setae’ on gecko toes. Until recently gecko setae were assumed to be composed entirely of keratin, but analysis of footprints left behind by geckos walking on surfaces revealed that setae include various kinds of lipids. However, the arrangement and molecular-level behavior of lipids and keratin in the setae is still not known. In the present study we demonstrate, for the first time, the use of Nuclear Magnetic Resonance (NMR) spectroscopy techniques to …


Urine Proteomics Identifies Preeclampsia Subphenotypes, Michelle R. Axe 2015 Otterbein University

Urine Proteomics Identifies Preeclampsia Subphenotypes, Michelle R. Axe

Undergraduate Honors Thesis Projects

Preeclampsia is a perinatal complication of unknown etiology, resulting in 76,000 maternal deaths and 500,000 infant deaths annually1. The Nationwide Children’s Center for Perinatal Research was the first cohort to propose that preeclampsia is rooted in protein misfolding. We aimed to characterize the misfolded protein components of maternal urine samples and to determine if distinct molecular profiles exist for clinical preeclampsia phenotypes. A total of 86 maternal urine samples from Yale-New Haven Hospital were analyzed for this study, from six clinical preeclampsia subphenotypes- pregnant control (n=10), idiopathic proteinuria (n=15), mild preeclampsia (n=14), superimposed preeclampsia (n=12), typical severe preeclampsia …


The Effect Of Red Maple Leaf Toxicosis On Reduced Glutathione Levels In Equine Erythrocytes In Vitro, Madeline A. Rohl 2015 Otterbein University

The Effect Of Red Maple Leaf Toxicosis On Reduced Glutathione Levels In Equine Erythrocytes In Vitro, Madeline A. Rohl

Undergraduate Honors Thesis Projects

Red maple leaf toxicosis is an equine blood disorder resulting from the consumption of wilted red maple (Acer rubrum L.) leaves by horses. Compounds within the leaves of red maple have oxidative effects on equine erythrocytes and can cause hemolysis of erythrocytes, the conversion of hemoglobin to methemoglobin, and the production of Heinz bodies. Reduced glutathione is important in the protection of equine erythrocytes from these oxidative events; however, in the presence of red maple toxin, glutathione is rapidly oxidized and is unavailable. The objective of this study is to determine whether the presence of vitamin C alters levels …


Investigating The Initial Detection Stage Of Meiotic Silencing By Unpaired Dna In The Model Organism Neurospora Crassa, Pegan A. Sauls 2015 Illinois State University

Investigating The Initial Detection Stage Of Meiotic Silencing By Unpaired Dna In The Model Organism Neurospora Crassa, Pegan A. Sauls

Theses and Dissertations

In an attempt to neutralize transposable elements or retrovirus invasions Neurospora crassa will rely on one of its many genome defense mechanism, Meiotic Silencing by Unpaired DNA (MSUD). MSUD works in a two-step process that first detects unpaired sequences between homologous chromosomes followed by downstream silenced expression of the sequence. The ultimate silencing stage of MSUD is widely accepted to operate through an RNAi-like system. However, the mechanics of the detection step of MSUD remains elusive. The research presented attempts to elaborate on how the initial stage of MSUD occurs and its specifics. First, a genetic approach is utilized to …


Methods In Extracting Dna: Using Phenol-Chloroform On Formalin Fixed Human Brain Tissue, Stefanie Rigg, Nathaniel Kennedy 2015 Purdue University North Central

Methods In Extracting Dna: Using Phenol-Chloroform On Formalin Fixed Human Brain Tissue, Stefanie Rigg, Nathaniel Kennedy

Undergraduate Research Conference

Genetic markers of diseases can be studied post-mortem with the isolation of DNA from tissues. While protocols exist for DNA extraction from fresh and formalin-fixed-paraffin embedded tissue and formalin fixed hard tissues such as bone, there are no published methods for formalin fixed soft tissue such as brain. Extractions from formalin fixed soft tissues are more difficult due to the higher amount of cellular degradation that results from fixation. The development of efficient DNA extraction protocols for formalin fixed soft tissues will allow post-mortem study of genetic markers and aid in identification of genetic diseases. Existing techniques of phenol-chloroform extraction …


Reagent Pencils: A New Technique For Solvent-Free Deposition Of Reagents Onto Paper-Based Microfluidic Devices, Haydn T. Mitchell, Isabelle C. Noxon, Cory A. Chaplan, Samantha J. Carlton, Cheyenne H. Liu, Kirsten A. Ganaja, Nathaniel W. Martinez, Chad Immoos, Philip Costanzo, Andres W. Martinez 2015 California Polytechnic State University, San Luis Obispo

Reagent Pencils: A New Technique For Solvent-Free Deposition Of Reagents Onto Paper-Based Microfluidic Devices, Haydn T. Mitchell, Isabelle C. Noxon, Cory A. Chaplan, Samantha J. Carlton, Cheyenne H. Liu, Kirsten A. Ganaja, Nathaniel W. Martinez, Chad Immoos, Philip Costanzo, Andres W. Martinez

Chemistry and Biochemistry

Custom-made pencils containing reagents dispersed in a solid matrix were developed to enable rapid and solvent-free deposition of reagents onto membrane-based fluidic devices. The technique is as simple as drawing with the reagent pencils on a device. When aqueous samples are added to the device, the reagents dissolve from the pencil matrix and become available to react with analytes in the sample. Colorimetric glucose assays conducted on devices prepared using reagent pencils had comparable accuracy and precision to assays conducted on conventional devices prepared with reagents deposited from solution. Most importantly, sensitive reagents, such as enzymes, are stable in the …


An Active Site Loop Mutant Of A Zinc Metallo-Deubiquitinase Suggests The Importance Of Loop Dynamics To Catalysis, Amy N. Bueno 2015 Purdue University

An Active Site Loop Mutant Of A Zinc Metallo-Deubiquitinase Suggests The Importance Of Loop Dynamics To Catalysis, Amy N. Bueno

Open Access Theses

AMSH is a conserved zinc metalloprotease that functions with endosomal sorting complexes required for transport (ESCRT) machinery in order to down regulate and degrade cell-surface receptors. It has been shown to have high specificity toward Lys63-linked polyubiquitin chains, a signal used for receptor down regulation by ESCRT machinery. It has previously been found that Sst2, an AMSH orthologue from fission yeast, contains a flexible "loop" near the proximal ubiquitin binding site, composed of residues L402 and F403. The different mobility of the side chains of these residues contribute to the opening and closing conformation of the catalytic cleft which allow …


Support Needs Of Young Ovarian Cancer Survivors: A Pilot Study, Aurelie Chuong 2015 Purdue University

Support Needs Of Young Ovarian Cancer Survivors: A Pilot Study, Aurelie Chuong

Open Access Theses

Purpose. Young ovarian cancer survivor (18 years old or older but diagnosed before the age of 34 years old) support needs (i.e., physical and daily living, psychological, sexual, patient care and support, health system and information) were examined to guide the design and implementation of future young adult and adolescent cancer support groups and interventions. Method. This cross-sectional pilot study included 11 young ovarian cancer survivors. The Supportive Care Needs Survey Short Form 34 (SCNS-SF34) was used and open-ended questions were added. Results. The calculated standardized scores for the SCNS-SF34 survey results showed that sexuality needs were ranked highest (41.7 …


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