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Biochemical Study Of Engineered Fluorescent Proteins As Calcium Sensors And The Effect Of Calcium And Ph In Cell Reproduction And Protein Expression, Malcom Arturo Delgado

Chemistry Theses

Calcium plays important roles in both eukaryotic and prokaryotic cells. Its actions help to stabilize cell synthesis, growth and development. In this thesis, studies have been completed to determine effects of calcium and pH on bacterial cell growth and protein expression using the bacterial cell strain E.coli BL21(DE3). Our studies demonstrated the addition of calcium addition in the media does not affect growth but increases protein expression, while reducing the pH from 7 to 4 through the addition of 10mM EGTA in LB media inhibits both. Additionally, we report studies on the design, expression, and purification of fluorescent mCherry variants …

Inhibition Of The Calcium Plateau Following In Vitro Status Epilepticus Prevents The Development Of Spontaneous Recurrent Epileptiform Discharges, Nisha Nagarkatti

Theses and Dissertations

Status epilepticus (SE) is a major clinical emergency resulting in continuous seizure activity that can cause brain injury and many molecular and pathophysiologic changes leading to neuronal plasticity. The neuronal plasticity following SE-induced brain injury can initiate epileptogenesis and lead to the ultimate expression of acquired epilepsy (AE), characterized clinically by spontaneous, recurrent seizures. Epileptogenesis is the process wherein healthy brain tissue is transformed into hyperexcitable neuronal networks that produce AE. Understanding these alterations induced by brain injury is an important clinical challenge and can lend insight into possible new therapeutic targets to halt the development of AE. Currently there …

Calcium Homeostasis And Cone Signaling Are Regulated By Interactions Between Calcium Stores And Plasma Membrane Ion Channels., Tamas Szikra, Peter Barabas, Theodore M. Bartoletti, Wei Huang, Abram Akopian, Wallace B. Thoreson, David Krizaj

Journal Articles: Ophthalmology

Calcium is a messenger ion that controls all aspects of cone photoreceptor function, including synaptic release. The dynamic range of the cone output extends beyond the activation threshold for voltage-operated calcium entry, suggesting another calcium influx mechanism operates in cones hyperpolarized by light. We have used optical imaging and whole-cell voltage clamp to measure the contribution of store-operated Ca(2+) entry (SOCE) to Ca(2+) homeostasis and its role in regulation of neurotransmission at cone synapses. Mn(2+) quenching of Fura-2 revealed sustained divalent cation entry in hyperpolarized cones. Ca(2+) influx into cone inner segments was potentiated by hyperpolarization, facilitated by depletion of …

Cd19 Signaling Is Impaired In Murine Peritoneal And Splenic B-1 B Lymphocytes, Trivikram Dasu, Vishal Sindhava, Stephen H. Clarke, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

B-1 cells reside predominantly within the coelomic cavities, tonsils, Peyer's patches, spleen (a minor fraction – ∼5%) and are absent in the lymph nodes. They are the primary sources of natural IgM in the body. B-1 cells express polyreactive B cell receptors (BCRs) that cross react with self-antigens and are thus implicated in auto-immune disorders. Previously, we reported that peritoneal B-1 cells are deficient in CD19-mediated intracellular signals leading to Ca²⁺ mobilization. Here, we find that splenic B-1 cells, like peritoneal B-1 cells, are defective in Ca²⁺ release upon B cell activation by co-cross-linking BCR and CD19. In …

The Relationship Between Calcium Intake, Obesity, And Cardiovascular Disease Risk Factors: The Jackson Heart Study, Marjuyua Lartey-Rowser

Dissertations

Cardiovascular disease (CVD) is a major health risk in the United States. Major indicators of CVD risk include obesity, blood lipids, and blood pressure. Modifiable risk factors associated with CVD include body composition (body mass index and waist circumference), serum lipids, and blood pressure. Data suggest calcium intake may play a role in regulation of weight, serum lipids, and blood pressure. The purpose of this study was to assess relationships of dietary calcium intake with weight status, and cardiovascular disease risks in African American population participating in the Jackson Heart Study.

The subjects included 4,267 African American adults ages 21-95 …

Differences In Response To “No Bones About It!” Between Boys And Girls, Amy Conklin

CFS Honors Program Undergraduate Theses

Early adolescence is a critical time for skeletal development, as this is when maximal bone growth occurs (1). Development of bones mass during adolescence can help reduce the risk for childhood fractures and osteoporosis later in life. Low calcium intakes can contribute to poor bone health and can increase the risk of developing osteoporosis (2). An adequate intake of 1300 milligrams per day of calcium was set for both male and female adolescents by the Institute of Medicine (3). Unfortunately, most adolescents do not attain this recommendation. Thus, researchers have aimed to investigate various effectively reaching young people to adopt …

Ionic Selectivity In L-Type Calcium Channels By Electrostatics And Hard-Core Repulsion, Dezso Boda, Douglas Henderson, Monika Valisko, Bob Eisenberg, Dirk Gillespie

Faculty Publications

A physical model of selective "ion binding" in the L-type calcium channel is constructed, and consequences of the model are compared with experimental data. This reduced model treats only ions and the carboxylate oxygens of the EEEE locus explicitly and restricts interactions to hard-core repulsion and ion–ion and ion–dielectric electrostatic forces. The structural atoms provide a flexible environment for passing cations, thus resulting in a self-organized induced-fit model of the selectivity filter. Experimental conditions involving binary mixtures of alkali and/or alkaline earth metal ions are computed using equilibrium Monte Carlo simulations in the grand canonical ensemble. The model pore rejects …

Zebrafish Kidney Phagocytes Utilize Macropinocytosis And Ca+-Dependent Endocytic Mechanisms., Claudia Hohn, Sang-Ryul Lee, Lesya M. Pinchuk, Lora Petrie-Hanson

College of Veterinary Medicine Publications and Scholarship

BACKGROUND: The innate immune response constitutes the first line of defense against invading pathogens and consists of a variety of immune defense mechanisms including active endocytosis by macrophages and granulocytes. Endocytosis can be used as a reliable measure of selective and non-selective mechanisms of antigen uptake in the early phase of an immune response. Numerous assays have been developed to measure this response in a variety of mammalian and fish species. The small size of the zebrafish has prevented the large-scale collection of monocytes/macrophages and granulocytes for these endocytic assays. METHODOLOGY/PRINCIPAL FINDINGS: Pooled zebrafish kidney hematopoietic tissues were used as …

Regulation Of Transglutaminase By 5-Ht2a Receptor Signaling And Calmodulin, Ying Dai

Dissertations

Transglutaminase (TGase), nature’s biological glue, catalyzes the post-translational modification of proteins by formation of intra- and intermolecular protein cross-links or by primary amine incorporation. TGase has various physiological functions, such as skin-barrier formation and blood clot stabilization, whereas increasing evidence indicates they may also involved in neurodegenerative diseases including Huntington’s disease (HD), Alzheimer disease’s, and progressive supranuclear palsy. Mutant huntingtin (htt) and small G proteins (e.g. Rac 1) are potential substrates of TGases. The purpose of this dissertation was to characterize the mechanisms by which 5-HT2A receptor signaling and calmodulin (CaM) regulate TGase-catalyzed transamidation of Rac1 and htt in cultured …

Furthering Pharmacological And Physiological Assessment Of The Glutamatergic Receptors At The Drosophila Neuromuscular Junction, Junyoung Lee

University of Kentucky Master's Theses

Drosophila larval neuromuscular junctions (NMJs) serve as a model for synaptic physiology. The molecular sequence of the postsynaptic glutamate receptors has been described; however, the pharmacological profile has not been fully elucidated. Despite the postsynaptic molecular sequence used to classify the receptors as a kainate subtype, they do not respond pharmacologically as such. Kainate does not depolarize the muscle, but dampens evoked EPSP amplitudes. Quantal responses show a decreased amplitude and area under the voltage curve indicative of reduced postsynaptic receptor sensitivity to glutamate transmission. ATPA, a kainate receptor agonist, did not mimic kainate’s action. The metabotropic glutamate receptor agonist …

Influence Of Calcium And Phosphorus Feeding On Markers Of Bone Metabolism In Transition Cows, V. R. Moreira, L. K. Zeringue, C. C. Williams, C. Leonardi, M. E. Mccormick

Faculty Publications

A study was carried out to verify the effect of Ca and P levels on production, digestibility, and serum bone metabolism biomarkers in dairy cows. Fifty-two nonlactating multiparous cows (≥3 lactations) were confined in a free-stall barn approximately 20 d before calving. A standard close-up diet was fed to cows once daily until d 2 postpartum. Cows were randomly assigned to 1 of 4 dietary treatments arranged in a 2 × 2 factorial approach averaging 0.64% Ca for high Ca (HCa), 0.46% Ca for low Ca (LCa), 0.47% P for high P (HP), and 0.38% P for low P (LP) …

The Effect Of Zinc On Cytokine Release And Signal Transduction In Airway Epithelial Cells, Nai-Lin Cheng

All ETDs from UAB

Zinc is an essential cation for life that is involved in numerous physiology and pathophysiology processes. Despite its wide use as a cytoprotective agent in medications, its cellular and molecular mechanisms of action have not been well established. In order to define the potential therapeutic benefit of zinc, I used several Cystic Fibrosis (CF) and non-CF human airway epithelial cell lines as models in my studies. Extracellular zinc caused release calcium from the endoplasmic reticulum though zinc-sensitive Gq-coupled receptor(s) that may subsequently modulate ion channels. In addition, zinc also exhibited anti-inflammatory effects. Zinc inhibited pro-inflammatory molecular TNFα-induced cytokine and chemokine …

Characterization Of Nprc And Its Binding Partners, Abdel A. Alli

USF Tampa Graduate Theses and Dissertations

The C type natriuretic peptide receptor (NPRC) also known as NPR3 is a widely expressed single transmembrane-spanning protein. NPRC functions as a homodimer at the cell surface for the metabolic clearance of a broad range of natriuretic peptides from circulation. The intracellular domain of NPRC is coupled to inhibitory G proteins and is involved in mediating signal transduction. In order to further elucidate the role of NPRC in signal transduction a proteomic approach was taken to identify putative protein binding partners for NPRC in different cell-types. An interrogation of the molecular association between NPRC and its identified protein binding partner(s) …

Correlation Of Calcium And Magnesium Intakes To Frequency Of Muscle Cramps In Female College Athletes, Jennifer Schneider, Marjorie Fitch-Hilgenberg

Discovery, The Student Journal of Dale Bumpers College of Agricultural, Food and Life Sciences

Muscle cramps are involuntary, painful, sudden contractions of skeletal muscles that can cause detrimental effects on athletic performance. Recent data suggest that low intakes of dietary calcium (Ca) and magnesium (Mg) can enhance or cause muscle cramps. The purpose of this study was to determine the correlation between Ca and Mg intakes and muscle cramping in female college athletes. Athletes completed a 24-hour dietary recall and a survey on frequency and location of muscle cramps. Of those surveyed, 8 participated in basketball, 21 in softball, and 10 in gymnastics. Calcium and Mg intakes were calculated from dietary recall data using …

In Vivo Function Of Otopetrin 1 In The Vestibular Sensory Epithelium, Euysoo Kim

All Theses and Dissertations (ETDs)

Otopetrin family genes encode multi-transmembrane domain proteins with three highly conserved domains. In mice, three Otopetrin paralogues are found. One of its members, Otopetrin 1: Otop1) has been previously shown to be essential for the formation of otoconia in the vestibular system of the inner ear. Otoconia are calcium carbonate biominerals that are required for normal balance and the sensation of linear acceleration with respect to gravity. The mechanism by which OTOP1 mediates otoconia biosynthesis is not known, but the ability of OTOP1 to modulate [Ca2+]i in response to purinergic signals in heterologous systems suggest that OTOP1 may be involved …

Primary Cilia: Highly Sophisticated Biological Sensors, Wissam A. Aboualaiwi, Shao T. Lo, Surya M. Nauli

Pharmacy Faculty Articles and Research

Primary cilia, thin hair-like structures protruding from the apical surface of most mammalian cells, have gained the attention of many researchers over the past decade. Primary cilia are microtubule-filled sensory organelles that are enclosed within the ciliary membrane. They originate at the cell surface from the mother centriole that becomes the mature basal body. In this review, we will discuss recent literatures on the roles of cilia as sophisticated sensory organelles. With particular emphasis on vascular endothelia and renal epithelia, the mechanosensory role of cilia in sensing fluid shear stress will be discussed. Also highlighted is the ciliary involvement in …

Structural Plasticity Can Produce Metaplasticity, Georgios Kalantzis, Harel Z Shouval

Journal Articles

BACKGROUND: Synaptic plasticity underlies many aspect of learning memory and development. The properties of synaptic plasticity can change as a function of previous plasticity and previous activation of synapses, a phenomenon called metaplasticity. Synaptic plasticity not only changes the functional connectivity between neurons but in some cases produces a structural change in synaptic spines; a change thought to form a basis for this observed plasticity. Here we examine to what extent structural plasticity of spines can be a cause for metaplasticity. This study is motivated by the observation that structural changes in spines are likely to affect the calcium dynamics …

Comment On "Generation Of Cold Low Divergent Atomic Beam Of Indium By Laser Ablation", A. Denning, A. Booth, S. Lee, M. Amonson, Scott D. Bergeson

Faculty Publications

We present measurements of the velocity distribution of calcium atoms in an atomic beam generated using a dual-stage laser back-ablation apparatus. Distributions are measured using a velocity selective Doppler time-of-flight technique. They are Boltzmann-like with rms velocities corresponding to temperatures above the melting point for calcium. Contrary to a recent report in the literature, this method does not generate a subthermal atomic beam.

The Interactive Effects Of Phosporus And Calcium On Phosphorus Requirement, Utilization, And Phytase Efficacy In Poultry, Syrena Powell

LSU Doctoral Dissertations

This research was conducted with broilers to investigate the interactive effects of P and Ca on P requirement, digestibility, utilization, and phytase efficacy. Each treatment had a minimum of 6 replications. One experiment with 3 trials was conducted to identify the nonphytate P (nPP) requirement of 0 to 14 d old broilers. The nPP levels fed ranged from 0.30 to 0.60% with a Ca:nPP of 2.22:1 or 1.9:1. Requirement estimates for 0 to 14 d ranged from 0.52 to 0.60% nPP. One experiment with 2 trials was conducted to identify the nPP requirement of 14 to 28 d old broilers. …