Biochemistry, Biophysics, and Structural Biology Commons^™

Biophysical Investigation Of Amyloid Formation And Their Prion-Like Self-Replication, Mentor Mulaj

USF Tampa Graduate Theses and Dissertations

Growth and deposition of amyloid fibrils, polymers of proteins with a cross beta-sheet structure, are associated with a significant number of human pathologies including Alzheimer’s disease, Parkinson’s disease, prion diseases, type II diabetes, and senile systematic or dialysis-related amyloidoses. The broader objective of my research is to identify the basic mechanisms regulating nucleation and growth of amyloid fibrils. There is increasing evidence that amyloid formation may proceed along at least two distinct assembly pathways for the formation of rigid fibrils. One pathway involves the nucleated polymerization of the characteristic rigid fibrils from partially denatured monomers and the other proceeds via …

The Interaction Between Eukaryotic Translation Initiation Factor Eif4g And 3’ Cap Independent Translation Element Of Barley Yellow Dwarf Virus Is Affected By Multiple Initiation Factors, Pei Zhao

Dissertations, Theses, and Capstone Projects

Barley Yellow Dwarf Virus (BYDV) lacks a 5’ (7-methyl guanosine) cap as well as a 3’poly A tail. Like many plant viruses, BYDV contains a cap independent translation element (CITE) in the 3’ untranslated region of the viral mRNA. BTE (Barley Yellow Dwarf Virus like cap-independent translation element) is one of the well characterized CITEs. BTE mediated translation primarily depends on eukaryotic initiation factor eIF4G. BTE binds to eIF4G; however, the details of BTE initiated translation are still unclear. Three eIF4G deletion mutants with different domain organization were used to investigate BTE interaction with eIF4G: eIF4G601-1196 is the eIF4G fragment …

Frontal Plane Comparison Between Drop Jump And Vertical Jump: Implications For The Assessment Of Acl Risk Of Injury, Guilherme Manna Cesar, Curtis L. Tomasevicz, Judith M. Burnfield

Athletic Performance Research

The potential to use the vertical jump (VJ) to assess both athletic performance and risk of anterior cruciate ligament (ACL) injury could have widespread clinical implications since VJ is broadly used in high school, university, and professional sport settings. Although drop jump (DJ) and VJ observationally exhibit similar lower extremity mechanics, the extent to which VJ can also be used as screening tool for ACL injury risk has not been assessed. This study evaluated whether individuals exhibit similar knee joint frontal plane kinematic and kinetic patterns when performing VJs compared with DJs. Twenty-eight female collegiate athletes performed DJs and VJs. …

Weight Control Practices Of Division I National Collegiate Athletic Association Athletes, Patrick B. Wilson, Leilani Madrigal, Judith M. Burnfield

Athletic Performance Research

Objectives: Altering body weight can have substantial effects on an athlete’s performance and well-being. Limited information is available describing the weight control practices of Division I National Collegiate Athletic Association (NCAA) athletes.

Methods: Weight control practices data from 188 (138 male and 50 female; 18-23 y) Division I NCAA athletes were analyzed as a part of this cross-sectional, retrospective study. Participants completed questionnaires on weight control practices and weight control nutrition knowledge at the end of their season and were classified into weight-sensitive and less weight-sensitive sports.

Results: A higher proportion of females attempted to lose weight than males among …

Heart And Sole: The Functional Role Of Fast-Skeletal Myosin Binding Protein-C In Cardiac And Skeletal Muscle, Brian Leei Lin

Dissertations

The goal of my dissertation was to compare and contrast the function of all three major isoforms of Myosin Binding Protein-C (MyBP-C): slow-skeletal, fast-skeletal, and cardiac (ssMyBP-C, fsMyBP-C, and cMyBP-C, respectively), with a focus on the least characterized isoform, fsMyBP-C. Using a variety of ex vivo, in vitro, and in silico methods, my research demonstrated that the N-terminal region of all MyBP-C isoforms bind to actin and shift tropomyosin, thus activating the thin filament during contraction. Furthermore, each isoform differentially activated the thin filament over isoform-specific ranges of Ca2+: slow-skeletal activates at low Ca2+, fast-skeletal activates at higher Ca2+, and …

A Mechanistic Study Of An Ipsc Model For Leigh’S Disease Caused By Mtdna Mutataion (8993 T>G), John P. Galdun

Theses and Dissertations

Mitochondrial diseases encompass a broad range of devastating disorders that typically affect tissues with high-energy requirements. These disorders have been difficult to diagnose and research because of the complexity of mitochondrial genetics, and the large variability seen among patient populations. We have devised and carried out a mechanistic study to generate a cell based model for Leigh’s disease caused by mitochondrial DNA mutation 8993 T>G. Leigh’s disease is a multi-organ system disorder that depends heavily on the mutation burden seen within various tissues. Using new reprogramming and sequencing technologies, we were able to show that Leigh’s disease patient fibroblasts …

Functional Effects Of Calcium Regulation Of Thin Filaments At Single Particle Resolution, Christopher Solis-Ocampo

Electronic Theses and Dissertations

Heart disease is the leading cause of death in the United States. Understanding heart function at the molecular level is critical for developing of more effective treatments. In the cardiac muscle, the thin filament is composed by troponin (Tn), tropomyosin (Tm), and F-actin. It provides Ca2+-dependent regulation of contraction by modulating myosin attachment and force generation in a cooperative scheme. However, this mechanism remains unclear. To understand thin filament activation, we studied the binding and functional properties of Tn and Tm to F-actin at single particle resolution by employing fluorescence image colocalization, in vitro motility assays, and Förster resonance energy …

A Dielectric Rod Antenna For Picosecond Pulse Stimulation Of Neurological Tissue, Ross A. Petrella, Karl H. Schoenbach, Shu Xiao

Bioelectrics Publications

A dielectrically loaded wideband rod antenna has been studied as a pulse delivery system to subcutaneous tissues. Simulation results applying 100-ps electrical pulse show that it allows us to generate a critical electric field for biological effects, such as brain stimulation, in the range of several centimeters. In order to reach the critical electric field for biological effects, which is similar to 20 kV/cm, at a depth of 2 cm, the input voltage needs to be 175 kV. The electric field spot size in the brain at this position is similar to 1 cm(2). Experimental studies in free space with …

Bayesian Signal Detection And Source Separation In Simulated Brain Computer Interface Systems, Muhammad Asim Mubeen

Legacy Theses & Dissertations (2009 - 2024)

The problems of signal detection and source separation are important in many fields of science and engineering. In many cases, a target signal needs to be detected in real time and is contaminated by noise. Sometimes the level of noise is on the order of the signal itself. The real time detection of a target signal is of key importance in problems such as the brain computer interface systems. In brain computer interface systems, the neural activity (electric signals) of the brain is detected using sensors (electrodes) on the surface of the brain or the scalp. This signal is contaminated …

Effects Of High Voltage Nanosecond Electric Pulses On Eukaryotic Cells (In Vitro): A Systematic Review, Tina Batista Napotink, Matej Reberšek, P. Thomas Vernier, Babara Mali, Damijan Miklavčič

Bioelectrics Publications

For this systematic review, 203 published reports on effects of electroporation using nanosecond high-voltage electric pulses (nsEP) on eukaryotic cells (human, animal, plant) in vitro were analyzed. A field synopsis summarizes current published data in the field with respect to publication year, cell types, exposure configuration, and pulse duration. Published data were analyzed for effects observed in eight main target areas (plasma membrane, intracellular, apoptosis, calcium level and distribution, survival, nucleus, mitochondria, stress) and an additional 107 detailed outcomes. We statistically analyzed effects of nsEP with respect to three pulse duration groups: A: 1–10 ns, B: 11–100 ns and C: …

The Effects Of The N-Terminal Extensions Of Cardiac Troponins On The Ca2+ Regulation Of Myosin Atpase Kinetics In Cardiac Myofibrils, Laura Gunther

Wayne State University Dissertations

Contraction of cardiac muscle is the basis of heart function. Heart failure, i.e., weakened contraction of cardiac muscle is the most common cause of morbidity and mortality of heart diseases. Cardiac muscle contraction is regulated by calcium via the function of troponin, a protein complex associated with the myofilaments in muscle cells. The cardiac troponin subunits T (cTnT) and I (cTnI) have unique N-terminal extensions that can be selectively removed by restrictive proteolysis during cardiac adaptation to physiological and pathological stresses, indicating a role of these proteins in modulating cardiac contraction. This study aims to understand the effects of the …

Examining The Reliability, Validity And Factor Structure Of The Drs-15 With College Athletes, Leilani Madrigal, Diane L. Gill, Kent M. Eskridge

Athletic Performance Research

Hardiness, comprising feelings of commitment, control and challenge, is most frequently measured with the Dispositional Resiliency Scale (DRS), but little work has been done with the brief 15-item version. To examine the factor structure, reliability, validity, and item functioning for gender invariance of the 15-item DRS with 525 collegiate athletes from a wide range of sports. Convergent and divergent validity were examined through relationships with mental toughness, grit and competitive anxiety. Participants completed measures of mental toughness, optimism, grit, competitive anxiety, and the DRS-15. Confirmatory Factor Analysis revealed a poor fit for the three-factor hardiness model, and subsequent exploratory factor …

Lipid-Protein Interactions Via Fluorescence Quenching In Small Unilamellar Vesicles, Talia Kordahi

Williams Honors College, Honors Research Projects

Studying peptide interactions with model membranes have aided in understanding protein-lipid interactions more clearly. In this report, fluorescent small unilamellar vesicles (SUVs) of a known composition were made. Two peptides, RND1 and MARCKS, were also fluorescently labeled and added to a solution of SUVs. The interactions between these were observed via fluorescence quenching, read out with a spectrofluorimeter. In addition, two environments of buffer were tested: 1X PBS and 0.1X PBS. After obtaining data, it was concluded that MARCKS quenched the SUVs more than RND1. This could be due to the presence of more lysine amino acids in MARCKS, which …

Two-Photon Excitation Based Photochemistry And Neural Imaging, Kevin Andrew Hatch

Open Access Theses & Dissertations

Two-photon microscopy is a fluorescence imaging technique which provides distinct advantages in three-dimensional cellular and molecular imaging. The benefits of this technology may extend beyond imaging capabilities through exploitation of the quantum processes responsible for fluorescent events. This study utilized a two-photon microscope to investigate a synthetic photoreactive collagen peptidomimetic, which may serve as a potential material for tissue engineering using the techniques of two-photon photolysis and two-photon polymerization. The combination of these techniques could potentially be used to produce a scaffold for the vascularization of engineered three-dimensional tissues in vitro to address the current limitations of tissue engineering. Additionally, …

Calcineurin: From Activation To Inhibition, Erik C. Cook

Theses and Dissertations--Molecular and Cellular Biochemistry

Calcineurin is a Ser/Thr phosphatase whose function is implicated in critical physiological processes such as immune system activation, fetal heart development, and long-term depression in neurons. Calcineurin has been implicated in the progression of Alzheimer’s disease and cardiac hypertrophy. It is not well understood how calcineurin is activated on a molecular level by Ca²⁺ and its activating protein calmodulin. Previous data from our lab show that calmodulin interaction induces the folding of the intrinsically disordered regulatory domain of calcineurin in two discrete and distant regions into α-helical conformations and that this folding is critical for complete activation of calcineurin. …

Noninvasive Measurement Of Electrical Events Associated With A Single Chlorovirus Infection Of A Microalgal Cell, Seung-Woo Lee, Eun-Hee Lee, Gerhard Thiel, James L. Van Etten, Ravi Saraf

Department of Chemical and Biomolecular Engineering: Faculty Publications

Chlorovirus Paramecium bursaria chlorella virus 1 (PBCV-1) contains a viral-encoded K⁺ channel imbedded in its internal membrane, which triggers host plasma membrane depolarization during virus infection. This early stage of infection was monitored at high resolution by recording the cell membrane depolarization of a single Chlorella cell during infection by a single PBCV-1 particle. The measurement was achieved by depositing the cells onto a network of one-dimensional necklaces of Au nanoparticles, which spanned two electrodes 70 μm apart. The nanoparticle necklace array has been shown to behave as a single-electron device at room temperature. The resulting electrochemical field-effect transistor …

Probing Allosteric, Partial Inhibition Of Thrombin Using Novel Anticoagulants, Stephen S. Verespy Iii

Theses and Dissertations

Thrombin is the key protease that regulates hemostasis; the delicate balance between procoagulation and anticoagulation of blood. In clotting disorders, like deep vein thrombosis or pulmonary embolism, procoagulation is up-regulated, but propagation of clotting can be inhibited with drugs targeting the proteases involved, like thrombin. Such drugs however, have serious side effects (e.g., excessive bleeding) and some require monitoring during the course of treatment. The reason for these side effects is the mechanism by which the drugs’ act. The two major mechanisms are direct orthosteric and indirect allosteric inhibition, which will completely abolish the protease’s activity. Herein we sought an …

High Affinity Block Of Icl,Swell By Thiol-Reactive Small Molecules, Sung H. Park

Theses and Dissertations

Ebselen (Ebs) is considered as a glutathione peroxidase (GPx) mimetic and primarily thought to function by scavenging intracellular reactive oxygen species (ROS). Previous to our work, Deng et al. (2010a) demonstrated complete block of I_Cl,swell with 15 microM Ebs following endothelin-1 (ET-1) induced activation of the current in cardiomyocytes. This block was presumed to take effect mainly via the quenching of ROS. Nonetheless, our work with DI TNC1 astrocytes strongly emphasizes that Ebs might function by an alternative mechanism based on its kinetic profile in blocking I_Cl,swell. Our experiments showed that 45 nM Ebs can fully block …

A Biophysical Study Of The Ion Transport Mechanism In Uncoupling Protein 2 By Investigating The Role Of Lysine Residues In Its Matrix Network, Stephanie O. Uwumarenogie

Theses and Dissertations (Comprehensive)

ABSTRACT

Uncoupling protein 2 (UCP2) is one of five UCP homologues found in the inner mitochondrial membrane that transports protons from the intermembrane space to the mitochondrial matrix. In turn, the proton motive force is dissipated and less ATP is produced in the mitochondria. UCP2 is proposed to influence insulin secretion in type II diabetes, and decrease the amount of reactive oxygen species produced in the mitochondria, however the detailed mechanism of ion (proton and anions) transport in UCP2 and other UCP homologues are not fully understood. Sequence alignment analysis performed on proteins in the mitochondrial carrier family (MCF) including …

Glucose-Fructose Likely Improves Gastrointestinal Comfort And Endurance Running Performance Relative To Glucose-Only, P. B. Wilson, S. J, Ingraham

Athletic Performance Research

This study aimed to determine whether glucose-fructose (GF) ingestion, relative to glucose-only, would alter performance, metabolism, gastrointestinal (GI) symptoms, and psychological affect during prolonged running. On two occasions, 20 runners (14 men) completed a 120-min submaximal run followed by a 4-mile time trial (TT). Participants consumed glucose-only (G) or GF (1.2:1 ratio) beverages, which supplied ~1.3 g/min of carbohydrate. Substrate use, blood lactate, psychological affect [Feeling Scale (FS)], and GI distress were measured. Differences between conditions were assessed using magnitude-based inferential statistics. Participants completed the TT 1.9% (−1.9; −4.2, 0.4) faster with GF, representing a likely benefit. FS ratings were …

Misrepresentation Of Carbohydrate For Exercise: ‘It Is Time To Bust The Myth Of Physical Inactivity And Obesity: You Cannot Outrun A Bad Diet’, Patrick B. Wilson

Athletic Performance Research

The contention from Malhotra et al1 regarding the unimportance of carbohydrate for exercise is misguided. First, the reference they provide is an editorial2 that should not be viewed as high-quality evidence. Several oft-cited exercise studies showing benefits with high-fat diets simultaneously utilized carbohydrate restoration strategies,3 which involves “carbohydrate loading” after a high-fat diet. Thus, the evidence provided does not contradict the established belief that carbohydrate is an important fuel for endurance exercise. Second, the authors’ use of the term “carbohydrate loading” is misleading in this context. No public health organizations advocate a diet containing carbohydrate in amounts used in “carbohydrate …

Structural And Functional Studies Of The Papain-Like Protease 2 From Mouse Hepatitis Virus, Yafang Chen

Open Access Dissertations

Our goal is to establish a system to investigate how the deubiquitinating (DUB) and deISGylating activities of coronavirus (CoV) papain-like protease domains (PLPs) are involved in virus immune evasion. To this end, we chose PLP2 from mouse hepatitis virus (MHV) as our target of study because MHV has historically served as a model system for the study of CoVs, and it has undeniable advantage of ease in culturing in comparison to human coronaviruses.

It is reported here the expression and purification of a region of MHV nsp3 that contains the catalytic core of the PLP2 domain and its neighboring domains. …

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 …

Discrete Davydov's Soliton In Α-Helical Protein Molecule With Anharmonic Hydrogen Bond And Thermal Noise, Frank T. Ndjomatchoua, Clément Tchawoua, Joël D.T. Tchinang, Bruno P. Lerü, Henri E.Z. Tonnang

All Peer-Reviewed Publications

An improvement of the Davydov's model for energy transfer in α-helix protein is proposed. Using hyperbolic cosine potential type to model hydrogen bond potential, we have obtained from the adiabatic approximation, a discrete nonlinear Schrödinger equation with inverse hyperbolic sine nonlinearity. Using the two-dimensional discrete map approach, we have found a linearly stable intrinsic localized mode of the corresponding developed equation. Mobility of the intrinsic localized mode was achieved by moving the local impurity hypothesized as the anomalous band energy. The reactivity of the discrete breather under thermal noise was studied numerically with the Langevin's approach. It was shown that …

Case 62: Glycolysis, The Pentose Phosphate Pathway, And The Citric Acid Cycle In Tumorigenesis, Dr. Kathleen Cornely

Chemistry & Biochemistry Case Studies

Hypoxia-inducible factors are transcription factors that mediate the cellular response to hypoxia (low cellular oxygen concentration) by enhancing or suppressing the transcription of specific genes involved in metabolic pathways.

Feasibility Study Of Inversely-Optimized Intensity Modulated Grid-Therapy For Bulky Lung Tumors, M Macfarlane, J Lam, R Dar, E Yu, K Jordan, Jz Chen

Data and Test Instruments

Purpose/Objective(s): Grid therapy with megavoltage photon beams has been clinically shown to improve local control of bulky tumors. The purpose of this study is to develop intensity modulated grid therapy (IMGT) with multiple beams. We hypothesize that when compared to conventional single-field grid therapy, IMGT will provide lower spatially fractionated doses to the surrounding organs at risk (OAR) while increasing the minimum dose to the whole tumor volume and thus improve tumor control probability as supported by conventionalradiobiology. Materials/Methods: IMGT was developed with multiple photon beams using dynamic MLC segments and orthogonal blocks. Spatially fractionated fluence patterns for all beams …

Biophysical Characterization Of A De Novo Elastin, Kelly Nicole Greenland

Dissertations, Theses, and Capstone Projects

Natural human elastin is found in tissue such as the lungs, arteries, and skin. This protein is formed at birth with no mechanism present to repair or supplement the initial quantity formed. As a result, the functionality and durability of elastin's elasticity is critically important. To date, the mechanics of this ability to stretch and recoil is not fully understood. This study utilizes de novo protein design to create a small library of simplistic versions of elastin-like proteins, demonstrate the elastin-like proteins, maintain elastin's functionality, and inquire into its structure using solution nuclear magnetic resonance (NMR).

Elastin is formed from …

Photoacoustic Calorimetry Studies Of The Earliest Events In Horse Heart Cytochrome-C Folding, Tarah A. Word

USF Tampa Graduate Theses and Dissertations

The protein folding problem involves understanding how the tertiary structure of a protein is related to its primary structure. Hence, understanding the thermodynamics associated with the rate-limiting steps for the formation of the earliest events in folding is most crucial to understanding how proteins adopt native secondary and tertiary structures. In order to elucidate the mechanism and pattern of protein folding, an extensively studied protein, Cytochrome-c (Cc), was chosen as a folding system to obtain detailed time-resolved thermodynamic profiles for the earliest events in the protein folding process. Cytochrome-c is an ideal system for understanding the folding process for several …

The Energy Benefits Of The Pantograph Wing Mechanism In Flapping Flight: Case Study Of A Gull, Gene D. Sattler

Faculty Publications and Presentations

Bird wings generally contain a 4-bar pantograph mechanism in the forearm that enables the wrist joint to be actuated from the elbow joint thus reducing the number of wing muscles and hence reducing the wing inertia and inertial drag. In this paper we develop a theoretical model of inertial power for flapping flight to estimate the advantage of the 4-bar pantograph mechanism by comparing the inertial power required for the case where wrist muscles are present in the forearm with the case where wrist muscles are not present in the forearm. It is difficult to predict how wrist muscles would …

Elucidating The Role Of Hausp Ubiquitin Like Domains In The Catalytic Function Of Usp7, Anuj Patel, Nicole Davis, Andrew Mesecar

The Summer Undergraduate Research Fellowship (SURF) Symposium

Ubiquitin specific proteases (USPs) are a class of enzymes involved in myriad cellular processes. One USP of great interest due to its oncogenic properties is USP7. In normal conditions USP7 is closely regulated due to its responsibility for destabilizing the tumor suppressor, p53, through the deubiquitination of MDM2. In multiple myeloma cases, it appears the regulation of USP7 subsides, as it is largely overexpressed, leading to the inappropriate degradation of p53. Inhibition of USP7 could, therefore, prove a viable target for cancer therapy. A greater understanding of USP7’s function and structure can lead to more insight into how this enzyme …