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Articles 61 - 85 of 85
Full-Text Articles in Physiology
Cellular And Matrix Changes In Articular Cartilage Of The Disproportionate Micromelia Mouse Model Of Osteoarthritis, Crystal Noelle Smaldone
Cellular And Matrix Changes In Articular Cartilage Of The Disproportionate Micromelia Mouse Model Of Osteoarthritis, Crystal Noelle Smaldone
Theses and Dissertations
Osteoarthritis (OA) is a degenerative joint disease that affects more than 60% of Americans 65 and older. Because human subjects and samples are not readily available for research, animal models are an invaluable resource for the study of OA. Disproportionate micromelia (Dmm) is one such model that develops OA early in life due to a deletion in the c-propeptide of the Col2a1 gene. Light microscope analysis of the articular cartilage in Dmm has been completed, but is insufficient to show the cellular effects of the deletion mutation in Dmm in adequate detail. The present study explores the changes that occur …
Differential Expression And Functional Characterization Of Alpha3 Beta2 Neuronal Nicotinic Acetylcholine Receptors, John Hideo Mizukawa
Differential Expression And Functional Characterization Of Alpha3 Beta2 Neuronal Nicotinic Acetylcholine Receptors, John Hideo Mizukawa
Theses and Dissertations
Neuronal nicotinic acetylcholine receptors (nAChRs) are expressed in both the periperhal and central nervous systems, and are involved in pre-, post-, and non-synaptic control of neuronal activation. In the brain, these receptors play an important role in a variety of physiological processes such as cognition, development, learning, and memory formation. Malfunction of these receptors have been implicated in neurodegenerative diseases like Alzheimer's disease (AD), schizophrenia, and Parkinson's disease. To date, 17 different nAChR subunits, including α2-α7 and β2-β4, have been cloned that can form homo- and/or hetero-pentameric ionotropic receptors. The unique combinations of subunit pentamers manifest in distinct functional receptors. …
Investigating And Modeling Possible Mechanisms By Which Healthy Cell Membranes Become Resistant To Hydrolysis By Secretory Phospholipase A2, Jennifer Nelson
Investigating And Modeling Possible Mechanisms By Which Healthy Cell Membranes Become Resistant To Hydrolysis By Secretory Phospholipase A2, Jennifer Nelson
Theses and Dissertations
Secretory phospholipase A2 (sPLA2) behaves differently toward the membranes of healthy cells compared to those of damaged or dying cells. The enzyme catalyzes rapid and sustained hydrolysis of compromised cells consistent with a simple catalytic mechanism. In contrast, when healthy cells are incubated with sPLA2, they become resistant to hydrolytic attack as manifest by three unusual observations: First, hydrolysis is transient and represents only a small fraction of the total membrane phospholipid content. Second, subsequent addition of sPLA2 fails to generate additional product. Third, the apparent potency of the enzyme to cause the membrane to be refractory is much greater …
A Comparative Analysis Of Hydrolysis Kinetics By Spla2 Isoforms During Apoptosis In S49 Cells, Erin Dalene Olson
A Comparative Analysis Of Hydrolysis Kinetics By Spla2 Isoforms During Apoptosis In S49 Cells, Erin Dalene Olson
Theses and Dissertations
Secretory Phospholipase A2 (sPLA2) represents a diverse class of roughly 20 enzymes, 12 of which have been identified in humans. These isoforms can be distinguished based on their tissue distribution, structure, and regulation. These differences in structure between the isoforms lead to the question does the enzyme's ability to respond to physical changes in the membrane during apoptosis governed by structure. S49 cell apoptosis was initiated by treatment with either the glucocorticoid dexamethasone (6–48 h) or with the calcium ionophore, ionomycin. The rates of hydrolysis were compared with each treatment condition for various concentrations of snake venom and human groups …
The Effects Of A Resveratrol Derivative On Regulatory Behaviors And Reproductive Health In Male And Female Long-Evans Rats, Kimberly Michelle Fabick
The Effects Of A Resveratrol Derivative On Regulatory Behaviors And Reproductive Health In Male And Female Long-Evans Rats, Kimberly Michelle Fabick
Theses and Dissertations
Phytoestrogens are chemicals produced by plants that act like estrogens and have the ability to bind to the mammalian estrogen receptor system. The purpose of this study is to evaluate a new phytoestrogen analog called 4-acetoxy Resveratrol. Resveratrol is a phytoestrogen that has been found in the skin of grapes. Resveratrol has been shown to be able to bind to the estrogen receptors and has a similar molecular structure as estradiol. Resveratrol has been shown to have many positive health benefits such as improving cardiovascular health, serving as a neuroprotective agent, acting as an anti-inflammatory agent, working as an anti-cancer …
The Effects Of Prenatally Administered Phytoestrogens On The Reproductive And Behavioral Development Of Long-Evans Rats, Crystal Blake
The Effects Of Prenatally Administered Phytoestrogens On The Reproductive And Behavioral Development Of Long-Evans Rats, Crystal Blake
Theses and Dissertations
Equol is known to be a selective androgen modulator and has the ability to bind and inhibit 5-alpha dihydrotestosterone (5α-DHT). Equol is also a selective estrogen receptor modulator and is able to bind beta estrogen receptors with high affinity. As estrogen receptors are found in the hypothalamus, pituitary, and gonads, prenatally administered equol could affect the morphological and reproductive development of offspring. To test this hypothesis, during gestational days 14 to 20, forty-two pregnant Long-Evans rats were given one of six treatments: 1) no treatment, 2) injection with dimethyl sulfoxide (DMSO), 3) injection with 10 mg/kg equol, 4) injection with …
Mechanisms By Which Apoptotic Membranes Become Susceptible To Secretory Phospholipase A2, Rachel Williams Bailey
Mechanisms By Which Apoptotic Membranes Become Susceptible To Secretory Phospholipase A2, Rachel Williams Bailey
Theses and Dissertations
During apoptosis, changes occur in T-lymphocyte membranes that render them susceptible to hydrolysis by secretory phospholipase A2 (sPLA2). To study the relevant mechanisms, a simplified model of apoptosis using a calcium ionophore was first applied. Kinetic and flow cytometry experiments provided key observations regarding ionophore treatment: initial hydrolysis rate was elevated, total reaction product was increased four-fold, and adsorption of the enzyme to the membrane surface was unaltered. Analysis of these results suggested that susceptibility during calcium-induced apoptosis is limited by substrate availability rather than enzyme adsorption. Fluorescence experiments identified three membrane alterations that might affect substrate access to the …
Thermodynamic Evidence That Ganglioside-Mediated Insertion Of Botulinum A Into The Cholinergic Nerve Ending May Precede Endocytosis And Acidification: A Langmuir Film Study, Bradley Adam Strongin
Thermodynamic Evidence That Ganglioside-Mediated Insertion Of Botulinum A Into The Cholinergic Nerve Ending May Precede Endocytosis And Acidification: A Langmuir Film Study, Bradley Adam Strongin
Theses and Dissertations
Botulinum Neurotoxin (BoNT) is one of the most potent toxins known to human kind. The Atomic Force Microscope (AFM) was employed to investigate the conditions under which BoNT type A heavy chain would bind and/or insert into mica supported dipalmitoylphosphatidylcholine (DPPC) lipid bilayers. As an alternate technique, DPPC/GT1b or total ganglioside extract (80:20) monolayers of a Langmuir Blodgett (LB) Trough were adapted to be artificial membrane models for toxin insertion studies. We conclude that LB monolayer studies are a promising candidate for BoNT/A membrane insertion investigation. Botulinum neurotoxin serotype A insertions into the LB monolayers in the presence of BoNT/A …
Effects Of Endurance Training On The Ampk Response To Exercise., David Gerald Chesser
Effects Of Endurance Training On The Ampk Response To Exercise., David Gerald Chesser
Theses and Dissertations
Activation of AMP-activated protein kinase (AMPK) results in the upregulation of several intracellular systems which help to prepare a cell for a high energy challenge. The magnitude of the AMPK response to a 10 min bout of exercise has been found to decrease in red quadriceps (RQ) following training, while putative AMPK roles seem to be maintained; specifically, the biogenesis of mitochondria and higher levels of hexokinase II and glucose transporter 4 (GLUT4). If the AMPK response to exercise is responsible in part for these adaptations, how can they be maintained if the AMPK response is attenuated? The purpose of …
Maternal High-Salt Diet During Pregnancy In Sprague Dawley Rats Programs Exaggerated Stress-Induced Blood Pressure And Heart Rate Responses In Adult Female Offspring, Summer Hayes King
Maternal High-Salt Diet During Pregnancy In Sprague Dawley Rats Programs Exaggerated Stress-Induced Blood Pressure And Heart Rate Responses In Adult Female Offspring, Summer Hayes King
Theses and Dissertations
The prenatal environment has been shown to have lasting effects on cardiovascular health. In the present study, pregnant rats were fed a 0.7% NaCl normal salt (NS) diet or an 8% NaCl high salt (HS) diet throughout pregnancy. Adult offspring were fitted with radiotelemetry probes to continuously measure blood pressure and heart rate. Rats were placed in restraining cages to test for a programmed acute stress hyperresponsiveness. Offspring were challenged with HS diet for one week to determine if blood pressure salt sensitivity had been programmed by the prenatal HS diet. Animals were killed following resting and acute stress conditions, …
Amp-Activated Protein Kinase Kinase Activity And Phosphorylation Of Amp-Activated Protein Kinase In Contracting Muscle Of Sedentary And Endurance Trained Rats, Denise Hurst
Theses and Dissertations
This study was designed to examine activity of AMP-activated protein kinase kinase (AMPKK) and AMP-activated protein kinase (AMPK) in muscles from control (C) and endurance trained (T) rats. Rats were trained 5 days/wk, 2 hr/d for 8 wks at a final intensity of 32 m/min up a 15% grade with 30 second sprints at 52 m/min every 10 min. Gastrocnemius muscles were stimulated in situ in T and C rats for 5 min at frequencies of 0.4/sec and 1/sec. Gastrocnemius LKB1 protein, a putative component of the AMPKK complex (LKB1, STRAD, and MO25), increased approximately 2-fold in response to training. …
Regulation Of Adrenal Steroidogenesis By Interleukin-6, Stephen A. Mcilmoil
Regulation Of Adrenal Steroidogenesis By Interleukin-6, Stephen A. Mcilmoil
Theses and Dissertations
Cortisol and dehydroepiandrosterone (DHEA) are steroids produced by the zona fasciculata (ZF) and reticularis (ZR), respectively, of the adrenal cortex. Both steroids are upregulated in response to adrenocorticotropic hormone (ACTH). Cortisol is a glucorticoid that is important in the regulation of inflammation and metabolism. DHEA is an adrenal androgen important in fetal growth and puberty but tends to decrease gradually after puberty in both men and women. DHEA has various effects on metabolism and immune function including inhibiting the effects of cortisol on some tissues. During the acute phase of stress, cortisol and DHEA rise due to an increase in …
Regulation Of Lkb1-Strad-Mo25 Complex Expression And Activation Of Ampk In Skeletal Muscle By Thyroid Hormone, Devon Jack Branvold
Regulation Of Lkb1-Strad-Mo25 Complex Expression And Activation Of Ampk In Skeletal Muscle By Thyroid Hormone, Devon Jack Branvold
Theses and Dissertations
AMP-activated protein kinase (AMPK), a heterotrimeric protein which serves as a metabolic master switch in skeletal muscle, is a research target for the pharmaceutical treatment and prevention of type 2 diabetes. The expression of all of the isoforms of the subunits of AMPK and AMPK activity are increased in skeletal muscle tissue of hyperthyroid rats. Activity of AMPK is regulated by an upstream kinase (AMPKK). The LKB1-STRAD-MO25 complex is a major AMPKK in skeletal muscle. This experiment was designed to determine whether the increase in AMPK activity is accompanied by a thyroid hormone-induced increase in the expression of the LKB1-STRAD-MO25 …
The Role Of Stat And The Jak/Stat Pathway In Mediating The Effects Of Interleukin-6 On Star Expression, Janae Strickland
The Role Of Stat And The Jak/Stat Pathway In Mediating The Effects Of Interleukin-6 On Star Expression, Janae Strickland
Theses and Dissertations
Cortisol, a hormone produced by a hormone produced by the adrenal gland, is responsible for many regulatory functions in the body. Cortisol release is mediated by adrenocorticotrophic hormone, or ACTH, through the hypothalamus-pituitary-adrenal or HPA axis. This HPA axis is the major release pathway used during acute stress, during which the levels of ACTH parallel those of cortisol. However, in states of chronic stress, the level of ACTH drops dramatically, while cortisol remains high. This study focuses on the pathway of cortisol release during these chronic stress states, specifically examining the role of IL-6 with respect to STATs and the …
Expression And Function Of Alpha3 And Beta2 Neuronal Nicotinic Acetylcholine Receptor Subunits In Hek-293 Cells, Nathan W. Steinhafel
Expression And Function Of Alpha3 And Beta2 Neuronal Nicotinic Acetylcholine Receptor Subunits In Hek-293 Cells, Nathan W. Steinhafel
Theses and Dissertations
Single-cell real-time quantitative RT-PCR was used to characterize the mRNA expression of rat neuronal nicotinic acetylcholine receptor (nAChR) subunits α3 and β2 in CA1 hippocampus stratum radiatum and stratum oriens interneurons. α3β2 co-expression was detected in 43% of interneurons analyzed. The nAChR subtype α3β2 was transiently expressed in cells derived from the human embryonic kidney cell line 293 at mRNA levels found in the CA1. The functional properties of α3β2 in HEK-293 cells were characterized by whole-cell patch clamping using acetylcholine (ACh) as an agonist. The kinetics of α3β2 channels were further analyzed by altering the level of α3 DNA …
The Influence Of Membrane Lipid Order On Cell Shape And Microvesiculation In Human Erythrocytes, Laurie Jackson Gonzalez
The Influence Of Membrane Lipid Order On Cell Shape And Microvesiculation In Human Erythrocytes, Laurie Jackson Gonzalez
Theses and Dissertations
Exposure of human erythrocytes to elevated intracellular calcium causes alterations in cell shape and stimulates shedding of the cell membrane in the form of microvesicles. We hypothesized that both the shape transition and microvesiculation are influenced by microscopic membrane physical properties such as lipid order. To test this hypothesis, membrane properties were manipulated by varying the experimental temperature, membrane cholesterol content, and the internal ionic environment. Changes in membrane order were assessed using steady-state fluorescence spectroscopy with an environment-sensitive probe, laurdan. Our observations led us to the following conclusions: 1) the modest temperature dependence of membrane structure observed with laurdan …
Fgf4 And Wnt5a/Pcp Signaling Promote Limb Outgrowth By Polarizing Limb Mesenchyme, Keri Lynn Low
Fgf4 And Wnt5a/Pcp Signaling Promote Limb Outgrowth By Polarizing Limb Mesenchyme, Keri Lynn Low
Theses and Dissertations
The focus of this study was to elucidate the molecular and cellular mechanisms whereby fibroblast growth factors (FGFs) mediate outgrowth of the limb. Specifically, we examined the epistatic relationship between FGF and Wnt/Planar cell polarity (PCP) signaling in establishing cell polarity as a mechanism for outgrowth. By implanting beads into embryonic limbs and lateral plate mesoderm, we established that FGF activates Wnt5a in a gradient fashion. Once it was established that Wnt5a was expressed at the right time and place to turn on PCP signaling, we investigated the ability of Wnt5a to influence cell migration and/or cell polarity. Our analysis …
Construction Of A Col11a1 Transgene Vector, Cameron Mckell Beck
Construction Of A Col11a1 Transgene Vector, Cameron Mckell Beck
Theses and Dissertations
Background: Cartilage disorders affect millions of people in the United States alone, with effects ranging from poor skeletal development and joint pain to shortened lifespan and perinatal lethality. Many of these disorders have their root in defects of collagen, type XI collagen being among the most important. A mouse model of such a type XI collagen defect is the chondrodysplasia (cho) mutant. Mice homozygous for this null mutation in the Col11a1 gene do not express the α1 chain of type XI collagen. This results in a functional knockout of type XI collagen, leading to insufficient skeletal development and perinatal lethality. …
The Effects Of 3-Phosphoglycerate And Other Metabolites On The Activation Of Amp-Activated Protein Kinase By Lkb1/Strad/Mo25, William John Ellingson
The Effects Of 3-Phosphoglycerate And Other Metabolites On The Activation Of Amp-Activated Protein Kinase By Lkb1/Strad/Mo25, William John Ellingson
Theses and Dissertations
Skeletal muscle contraction results in the phosphorylation and activation of the AMP-activated protein kinase (AMPK) by an upstream kinase, AMPKK. The LKB1-STRAD-MO25 complex is the major AMPKK in skeletal muscle; however, LKB1-STRAD-MO25 activity is not increased by muscle contraction. This relationship suggests that phosphorylation of AMPK by LKB1-STRAD-MO25 during skeletal muscle contraction may be regulated by allosteric mechanisms. In this study we tested an array of metabolites including glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), fructose 1,6-bisphosphate (F1,6-P2), 3-phosphoglycerate (3PG), glucose-1-phosphate (G1P), glucose-1,6-bisphosphate (G1,6-P2), adenosine diphosphate (ADP), carnitine (Carn), acetyl-carnitine (Acarn), inosine monophosphate (IMP), inosine, and ammonia for allosteric regulation. We found that …
Pka As An Upstream Kinase For Lkb1/Strad/Mo25, Seth Taylor Herway
Pka As An Upstream Kinase For Lkb1/Strad/Mo25, Seth Taylor Herway
Theses and Dissertations
The LKB1/STRAD/MO25 complex (LSMK) has been identified as the major upstream kinase for AMP-activated protein kinase (AMPK). PKA phosphorylates LKB1 at the Ser428 residue in humans and Ser431 residue in mice. We investigated PKA as an upstream kinase for LSMK. LKB1 that had been incubated with PKA prior to incubation with AMPK experienced up to a 51% increase in AMPK Kinase activity compared to LKB1 alone (p < 0.05). When blocked with a PKA Inhibitor, the kinase effect of PKA on LKB1 was eliminated. Rat epitrochlearis muscle tissue incubated with epinephrine experienced no increase in AMPK activity compared with controls indicating that epinephrine does not cause AMPK activity in this type of tissue. In conclusion, phosphorylation by PKA can increase the AMPKK activity of LKB1-STRAD-MO25 in vitro. Because LKB1 has been found to be constitutively active, it is postulated that phosphorylation by PKA may act to enhance LKB1-AMPK interaction and thus achieve its effect.
Properties Of Conductance And Inhibition Of Proton Channels: M2 From Influenza A Virus And Fo From Escherichia Coli Atp Synthase, Jeffrey C. Moffat
Properties Of Conductance And Inhibition Of Proton Channels: M2 From Influenza A Virus And Fo From Escherichia Coli Atp Synthase, Jeffrey C. Moffat
Theses and Dissertations
Proton channels are essential for many of the processes of life. The influenza A viral protein M2 is responsible for sensing the conditions necessary for viral RNA release. The proton-translocating FoF1 ATPase (ATP synthase) uses a proton gradient to drive adenosine triphosphate (ATP) synthesis. We have directly measured proton uptake in vesicles containing reconstituted M2 or FO by monitoring external pH after addition of valinomycin to vesicles with 100-fold diluted external [K+]. This proton flux assay was utilized to quantify proton flux through single M2 and Fo channels. Contrary to previous reports, proton uptake by M2 was not significantly altered …
The Role Of Fgfr4 In Trigeminal Placode Cell Development, Stephanie Beth Reynolds
The Role Of Fgfr4 In Trigeminal Placode Cell Development, Stephanie Beth Reynolds
Theses and Dissertations
In vertebrate embryos, the ophthalmic trigeminal (opV) placode contributes sensory neurons to the trigeminal ganglion during development of the peripheral nervous system. FGFR4 is expressed transiently in the trigeminal placode as cells undergo an epithelial-mesenchymal transition and begin to migrate toward the condensing ganglion. Because of the restricted spatiotemporal expression of FGFR4 in the opV placode, it was hypothesized that FGFR4 is necessary for the process of epithelial-mesenchymal transition in the opV placode. Using electroporation, an FGFR4 inhibitory gene construct was introduced into 6—10 somite stage chick embryos. This secreted inhibitory form of the FGFR4 gene blocked endogenous FGFR4 signaling, …
The Role Of Pax3 In Neuronal Differentiation Of The Ophthalmic (Opv) Trigeminal Placode And Neural Tube During Chicken Embryonic Development, James R. Bradshaw
The Role Of Pax3 In Neuronal Differentiation Of The Ophthalmic (Opv) Trigeminal Placode And Neural Tube During Chicken Embryonic Development, James R. Bradshaw
Theses and Dissertations
Pax3 has been used as a valuable marker in research aimed at understanding tissue interactions involved in trigeminal ophthalmic (opV) placode development. While Pax3 expression coincides with opV neuron specification, the function of Pax3 in these cells has not previously been investigated. Splotch mutant mice (which lack Pax3) have a reduced trigeminal ganglion; however it is not clear whether this reduction is due to neural crest or placode cells. We have used electroporation in the chick model system to block or ectopically express Pax3 at key times in opV placode development. Using several markers of placode cell differentiation, we have …
Phosphorylation Of Skeletal Muscle Acetyl-Coa Carboxylase By Ampk Enhances Palmitoyl-Coa Inhibition, Dustin S. Rubink
Phosphorylation Of Skeletal Muscle Acetyl-Coa Carboxylase By Ampk Enhances Palmitoyl-Coa Inhibition, Dustin S. Rubink
Theses and Dissertations
Acetyl-CoA carboxylase (ACC) catalyzes the formation of malnoyl-CoA, which in turn controls the rate of fatty acid metabolism. ACC beta or 2 has been shown to be localized on the mitochondria in close proximity to carnintine palmitoyl transferase 1 (CPT-1), the enzyme responsible for the influx of acyl-CoA into the matrix where beta oxidation takes place. CPT-1 is inhibited by malonyl-CoA produced by ACC. It has been well documented that AMP activated kinase (AMPK) when activated phosphorylates and inactivates ACC. ACC is controlled allosterically by citrate, which activates, and by palmitoyl-COA, which inhibits. In this study, we asked the question, …
Mechanism Governing The Cellular Susceptibility To Secretory Phospholipase A2, Lauren Blackburn Jensen
Mechanism Governing The Cellular Susceptibility To Secretory Phospholipase A2, Lauren Blackburn Jensen
Theses and Dissertations
Secretory phospholipase A2 (sPLA2) is an important part of apoptosis and disposal of damaged and dying cells. However, healthy cells are not susceptible to attack by sPLA2. Recent studies have focused on membrane properties necessary to induce susceptibility in both artificial and biological membranes. Hydrolysis of phospholipids by sPLA2 requires at least two preliminary steps: first, adsorption of the enzyme to the cellular membrane, and second, movement of a phospholipid into the active site of the enzyme. We determined the effects of susceptibility on each of the two steps and determined the contributions changing the equilibrium constants have on susceptibility. …