Biochemistry, Biophysics, and Structural Biology Commons^™

Extracullular Atp Regulates Il-1beta Release From Microglial Cells Via Purinergic Receptor After In Vitro Trauma, Chengya Liang

Electronic Theses and Dissertations

Traumatic brain injury (TBI) induces a state of microglialactivation, which includes upregulation of macrophage functions and release inflammatory mediators such as certain inflammatory cytokines. Current literature suggests that interleukin-1Beta is an important cytokine mediator, which is dramatically increased after brain injury. Previous studies indicate that ATP is released by traumatically injured astrocytes and serves as a cell-to-cell mediator through purinergic receptors after in vitro injury. However, the mechanism of interleukin-1Beta release after traumatic brain injury remains poorly defined and is difficult to study using in vivo models. Using an in vitro model for traumatic brain injury (cell strain or stretch), …

Possible Use Of P20 Antigen In Serodiagnosis Of Inflammatory Bowel Disease, Shinchieh Yang

Electronic Theses and Dissertations

Crohn's disease (CD) is an idiopathic, chronic, relapsing inflammatory disorder, which is most commonly involved terminal ileum and colon. The incidence and prevalence of CD has dramatically increased during the last 50 years; however, the etiology and mechanism of this disorder remain unveiled. Besides genetic susceptibility, recent integrated researches investigated the role of environmental triggers such as microflora, measles viruses and mycobacteria in the pathogenesis of CD. The association between M. avium subsp paratuberculosis (MAP) and CD has been heightened because of clinical resemblance to Johne's disease (JD), a granulomatous enteritis in ruminants caused by MAP. Isolation of MAP from …

Evaluation Of Immunogenicity Of Transgenic Chloroplast Derived Protect, Vijay Koya

Electronic Theses and Dissertations

Anthrax, a fatal bacterial infection is caused by Bacillus anthracis, a gram-positive, spore forming, capsulated, rod shaped organism. Centers for Disease Control (CDC) lists anthrax as Category A biological agent due to its severity of impact on human health, high mortality rate, acuteness of the disease and potential for delivery as a biological weapon. The currently available human vaccine in the United States (AVA anthrax vaccine adsorbed) is prepared from Alum adsorbed formalin treated supernatant culture of toxigenic, non-encapsulated strain of Bacillus anthracis with the principle component being protective antigen (PA83). Evaluation of anthrax vaccine given to nearly 400,000 US …

Survival Of Mycobacterium Avium Subspecies Paratuberculosis In The Pol, John Rumsey

Electronic Theses and Dissertations

Mycobacterium avium subspecies paratuberculosis (map) is an intracellular pathogen that is known to parasitize macrophages and monocytes. Map infiltrates gastrointestinal tract host tissue where it is the known etiological agent of johne's disease in ruminants and implicated in the etiology of crohn's disease in humans. Map's ability to survive within macrophages enables it to disseminate throughout the rest of the host, possibly infecting other circulating blood leukocytes. In this study, the survival and fate of map strain atcc 43015 (human isolate) following phagocytosis was determined using in vitro murine macrophage cell line j774a.1 and polymorphonuclear cells (pmnc's) from five crohn's …

Characterization Of A Novel Interactor/Substrate For The Pro-Apoptotic Serine Protease Omi/Htra2, Valerie Anne Stratico

Electronic Theses and Dissertations

OmiHtrA2 is a highly conserved mammalian serine protease that belongs to the HtrA family of proteins. Omi shares homology with the bacterially expressed heat shock protease HtrA, which functions as a protease at higher temperatures and a chaperone at lower temperatures. Additionally, Omi shares sequence similarity with the mammalian homologs L56/HtrA1 and PRSP/HtrA3. Omi was first isolated as an interacting protein of Mxi2, an alternatively spliced form of the p38 stress-activated kinase, using a modified yeast two-hybrid system. Omi localizes in the mitochondria and in response to apoptotic stimuli the mature form of this protein translocates to the cytoplasm. In …

Molecular Typing Of Mycobacterial Isolates Cultured From The Tissue Of Inflammatory Bowel Disease (Crohn's Disease) Patients, Leanne M. Adams

Electronic Theses and Dissertations

The role of Mycobacterium avium subsp paratuberculosis (MAP) in the etiology and pathogenesis of inflammatory bowel disease (IBD) including Crohn's Disease (CD), has been investigated. The fastidious characteristics and cross reactivity of MAP with other members in Mycobacteria have produced significant challenges in their detection and identification. In this two year pilot study, an array of three PCR molecular assays based on the detection of sequences from the16S rRNA, IS1245, and IS900 genes, belonging to members of the MAC, have been developed and optimized into a common protocol to be used as a rapid and accurate diagnostic tool regarding M. …

A Role For Talin In Presynaptic Function, Jennifer R. Morgan, Gilbert Di Paolo, Hauke Werner, Valentina A. Shchedrina, Marc Pypaert, Vincent A. Pieribone, Pietro De Camilli

Department of Biochemistry: Faculty Publications

Talin, an adaptor between integrin and the actin cytoskeleton at sites of cell adhesion, was recently found to be present at neuronal synapses, where its function remains unknown. Talin interacts with phosphatidylinositol-(4)-phosphate 5-kinase type Iγ, the major phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P₂]–synthesizing enzyme in brain. To gain insight into the synaptic role of talin, we microinjected into the large lamprey axons reagents that compete the talin–PIP kinase interaction and then examined their effects on synaptic structure. A dramatic decrease of synaptic actin and an impairment of clathrin-mediated synaptic vesicle endocytosis were observed. The endocytic defect included an accumulation of clathrin-coated pits …

Methionine Sulfoxide Reductase Regulation Of Yeast Lifespan Reveals Reactive Oxygen Species-Dependent And -Independent Components Of Aging, Ahmet Koc, Audrey P. Gasch, Julian C. Rutherford, Hwa-Young Kim, Vadim N. Gladyshev

Department of Biochemistry: Faculty Publications

Aging is thought to be caused by the accumulation of damage, primarily from oxidative modifications of cellular components by reactive oxygen species (ROS). Here we used yeast methionine sulfoxide reductases MsrA and MsrB to address this hypothesis. In the presence of oxygen, these antioxidants could increase yeast lifespan and did so independent of the lifespan extension offered by caloric restriction. However, under ROS-deficient, strictly anaerobic conditions, yeast lifespan was shorter, not affected by MsrA or MsrB, and further reduced by caloric restriction. In addition, we identified changes in the global gene expression associated with aging in yeast, and they did …

The Identity Of Proteins Associated With A Small Heat Shock Protein During Heat Stress In Vivo Indicates That These Chaperones Protect A Wide Range Of Cellular Functions, Eman Basha, Garrett J. Lee, Linda A. Breci, Andrew C. Hausrath, Nicole R. Baun, Kim C. Giese, Elizabeth Vierling

Department of Biochemistry: Faculty Publications

The small heat shock proteins (sHSPs) are a ubiquitous

class of ATP-independent chaperones believed to

prevent irreversible protein aggregation and to facilitate

subsequent protein renaturation in cooperation

with ATP-dependent chaperones. Although sHSP chaperone

activity has been studied extensively in vitro, understanding

the mechanism of sHSP function requires

identification of proteins that are sHSP substrates in

vivo. We have used both immunoprecipitation and affinity

chromatography to recover 42 proteins that specifically

interact with Synechocystis Hsp16.6 in vivo during

heat treatment. These proteins can all be released from

Hsp16.6 by the ATP-dependent activity of DnaK and cochaperones

and are heat-labile. …

Development Of A Gene Transfer System In Clostridium Scindens Vpi 12708, Rashmi Ramasubbaiah

Masters Theses & Specialist Projects

Clostridium scindens VPI 12708 (previously known as Eubacterium sp. VPI 12708) is a bile-acid dehydroxylating bacterium originally isolated from the feces of a colon cancer patient. Many genes required for bile acid 7-a dehydroxylation are found on a large bile acid inducible operon (bai) that has been extensively studied. However the bai promoter, which directs expression of the bai operon, has yet to be characterized due, in part, to a lack of a functional genetic transfer system for this strain. A spontaneous rifampinresistant Clostridium scindens VPI 12708 mutant was used as a recipient to determine the efficacy of conjugation as …

Aminoacyl-Trnas: Setting The Limits Of The Genetic Code, Michael Ibba, Dieter Söll

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Aminoacyl-tRNAs (aa-tRNAs) are simple molecules with a single purpose—to serve as substrates for translation. They consist of mature tRNAs to which an amino acid has been esterified at the 3′-end. The 20 different types of aa-tRNA are made by the 20 different aminoacyl-tRNA synthetases (aaRSs, of which there are two classes), one for each amino acid of the genetic code (Ibba and Söll 2000). This would be fine if it were not for the fact that such a straightforward textbook scenario is not true in a single known living organism. aa-tRNAs lie at the heart of gene expression; they interpret …

Theoretical Neuroscience: Modeling The Activation Mechanism Of Potassium Channels In Neurons, Kevin Twedt

Presidential Scholars Theses (1990 – 2006)

We have modeled the electrostatic interaction between the S4 segment of the potassium channel molecule and the surrounding water molecules on both the intracellular and extracellular sides of the neural axon cell membrane. Two methods were used to approximate this interaction: (i) a macroscopic evaluation in which the water was treated as a dielectric medium with dielectric constant 80; (ii) a microscopic evaluation considering the effects of each individual water molecule fixed in position within the water pockets surrounding the S4 segment. The potential energy of the S4 due to the water pockets was plotted against the rotation of the …

Effects Of Overexpressing Alternative Isoform Pax-5e In Vivo, Suzanne Elizabeth Cole

Dissertations, Theses, and Masters Projects

No abstract provided.

Identification Of Phosphorylase Kinase Alpha Subunit Binding Partners In Skeletal Muscle, Soleil Archila

Masters Theses & Specialist Projects

IDENTIFICATION OF PHOSPHORYLASE KINASE ALPHA SUBUNIT BINDING PARTNERS IN SKELETAL MUSCLE Soleil Archila August 2004 67 Pages Directed by: Nancy A. Rice, Sigrid Jacobshagen, and Claire A. Rinehart Department of Biology Western Kentucky University Phosphorylase kinase (PhK) integrates neural, hormonal, and metabolic signals in skeletal muscle to tightly regulate glycogen breakdown and energy production. Structurally, PhK is among the largest and most complex kinases known with a stoichiometry of (αβγδ)4 and a mass of 1.3x106 Da. The catalytic γ subunit is allosterically controlled through alterations in quaternary structure initiated by the regulatory α, β and δ subunits. In this study …

Computational Protein Biomarker Prediction: A Case Study For Prostate Cancer, Michael Wagner, Dayanand N. Naik, Alex Pothen, Srinivas Kasukurti, Raghu Ram Devineni, Bao-Ling Adam, O. John Semmes, George L. Wright Jr.

Mathematics & Statistics Faculty Publications

Background: Recent technological advances in mass spectrometry pose challenges in computational mathematics and statistics to process the mass spectral data into predictive models with clinical and biological significance. We discuss several classification-based approaches to finding protein biomarker candidates using protein profiles obtained via mass spectrometry, and we assess their statistical significance. Our overall goal is to implicate peaks that have a high likelihood of being biologically linked to a given disease state, and thus to narrow the search for biomarker candidates.

Results: Thorough cross-validation studies and randomization tests are performed on a prostate cancer dataset with over 300 patients, obtained …

Dissecting The Loci Of Low-Level Quinine Resistance In Malaria Parasites, Michael T. Ferdig, Roland A. Cooper, Jianbing Mu, Bingbing Deng, Deirdre A. Joy, Xin-Zhuan Su, Thomas E. Wellems

Biological Sciences Faculty Publications

Quinine (QN) remains effective against Plasmodium falciparum, but its decreasing efficacy is documented from different continents. Multiple genes are likely to contribute to the evolution of QN resistance. To locate genes contributing to QN response variation, we have searched a P. falciparum genetic cross for quantitative trait loci (QTL). Results identify additive QTL in segments of chromosomes (Chrs) 13, 7 and 5, and pairwise effects from two additional loci of Chrs 9 and 6 that interact, respectively, with the QTL of Chrs 13 and 7. The mapped segments of Chrs 7 and 5 contain pfcrt, the determinant of …

Effects Of Sham Air And Cigarette Smoke On A549 Lung Cells: Implications For Iron-Mediated Oxidative Damage, Jonathan J. Mayo, Pete Kohlhepp, Dianzheng Zhang, Joy J. Winzerling

PCOM Scholarly Papers

Inhalation of airborne pollution particles that contain iron can result in a variety of detrimental changes to lung cells and tissues. The lung iron burden can be substantially increased by exposure to cigarette smoke, and cigarette smoke contains iron particulates, as well as several environmental toxins, that could influence intracellular iron status. We are interested in the effects of environmental contaminants on intracellular iron metabolism. We initiated our studies using lung A549 type II epithelial cells as a model, and we evaluated the effects of iron dose and smoke treatment on several parameters of intracellular iron metabolism. We show that …

Activation Kinetics Of Skinned Cardiac Muscle By Laser Photolysis Of Nitrophenyl-Egta, Hunter Martin, Marcus G. Bell, Graham C. Ellis-Davis, Robert J. Barsotti

PCOM Scholarly Papers

The kinetics of Ca2+-induced contractions of chemically skinned guinea pig trabeculae was studied using laser photolysis of NP-EGTA. The amount of free Ca2+ released was altered by varying the output from a frequency-doubled ruby laser focused on the trabeculae, while maintaining constant total [NP-EGTA] and [Ca2+]. The time courses of the rise in stiffness and tension were biexponential at 23°C, Ph 7.1, and 200 Mm ionic strength. At full activation (pCa < 5.0), the rates of the rapid phase of the stiffness and tension rise were 56 ± 7 s-1 (n = 7) and 48 ± 6 s-1 (n = 11) while the amplitudes were 21 ± 2 and 23 ± 3%, respectively. These rates had similar dependencies on final [Ca2+] achieved by photolysis: 43 and 50 s-1 per Pca unit, respectively, over a range of [Ca2+] producing from 15% to 90% of maximal isometric tension. At all [Ca2+], the rise in stiffness initially was faster than that of tension. The maximal rates for the slower components of the rise in stiffness and tension were 4.1 ± 0.8 and 6.2 ± 1.0 s-1. The rate of this slower phase exhibited significantly less Ca2+ sensitivity, 1 and 4 s-1 per Pca unit for stiffness and tension, respectively. These data, along with previous studies indicating that the force-generating step in the cross-bridge cycle of cardiac muscle is marginally sensitive to [Ca2+], suggest a mechanism of regulation in which Ca2+ controls the attachment step in the cross-bridge cycle via a rapid equilibrium with the thin filament activation state. Myosin kinetics sets the time course for the rise in stiffness and force generation with the biexponential nature of the mechanical responses to steps in [Ca2+] arising from a shift to slower cross-bridge kinetics as the number of strongly bound cross-bridges increases.

Regulation Of Myocardial Hsp70 Gene Expression Following Exercise, C.W. Melling, David Thorp, Earl Noble

Jamie Melling

No abstract provided.

The Role Of Amylomaltase In Maltose Metabolism In The Cytosol Of Photosynthetic Cells, Yan Lu, Thomas Sharkey

Yan Lu

No abstract provided.

Intravenous 2-Deoxy-D-Glucose Injection Rapidly Elevates Levels Of The Phosphorylated Forms Of P44/42 Mitogen-Activated Protein Kinases (Extracellularly Regulated Kinases 1/2) In Rat Hypothalamic Parvicellular Paraventricular Neurons, Arshad Khan, Alan G. Watts

Arshad M. Khan, Ph.D.

No abstract provided.

Innovation Education In Nsw Design And Technology Curriculum, A Turner, K Seemann

Dr Angela Turner

Do technology teachers in secondary schools and academic stakeholders share the national vision of knowledge and innovation when implementing Design and Technology curricula? Directions for an innovation climate have been endorsed by the federal government, and demanded by various industry groups, since 1996. This paper explores the extent to which education providers of secondary schooling have embraced the call for teaching and developing innovation capacities through technology curriculum. The Australian Science, Technology and Engineering Council (ASTEC) in 1996 underpinned “foresight” as an essential dimension to our thinking which attempts to capture the dynamics of change and the need to incorporate …