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Articles 1 - 26 of 26
Full-Text Articles in Physiology
Systemic And Microcirculatory Effects Of Hboc-201: A Hemoglobin-Based Oxygen Carrier, Bjorn Song
Systemic And Microcirculatory Effects Of Hboc-201: A Hemoglobin-Based Oxygen Carrier, Bjorn Song
Theses and Dissertations
Top-loading This study compared the effects on oxygen (O2) transport of four fluids: 5.9% human serum albumin (HSA) a non-O2 carrying iso-oncotic solution (volume control);HBOC-201 (Hemopure, Biopure Corp., Cambridge, MA); MP50, HBOC-201 with a P50 of 18 mmHg; and lastly LP50, an HBOC-201 with P50 of 17 mmHg and higher viscosity (4 cP). It has been proposed that HBOC with a higher O2 affinity and a viscosity closer to that of whole blood will cause less vasoconstriction and thus a lower MAP than Hemopure (P50 = 40 mmHg and 2.2 cP). Intravital microscopic measurements were made on the spinotrapezius muscle …
Proteoliposome Proton Flux Assays Establish Net Conductance, Ph-Sensitivity, And Functional Integrity Of A Novel Truncate Of The M2 Ion "Channel" Of Influenza A, Emily Peterson
Theses and Dissertations
A novel truncate of Influenza A M2 protein (residues 22-62), incorporated into a uniquely tailored proteoliposome proton uptake assay, demonstrated proton flux more characteristic of an ion transporter than a traditional ion "channel." The liposome paradigm was essential for testing the conductance activity of this M2 truncate at a range of extraphysiological pHs appropriate for channel vs. transport function determination. In addition to transporter-typical proton flux, M2(22-62) showed the key characteristics of functional integrity: selective proton uptake into liposomes and block of uptake by amantadine. Two sets of proteoliposome proton flux assays were carried out, Set 1 at pH values …
Characterization Of The Lkb1-Mo25-Strad Ampkk Complex In Adult Mouse Skeletal Muscle, Cody Don Smith
Characterization Of The Lkb1-Mo25-Strad Ampkk Complex In Adult Mouse Skeletal Muscle, Cody Don Smith
Theses and Dissertations
In liver tissue, the AMP-activated protein kinase kinase (AMPKK) complex was identified as the association of LKB1, MO25α/β, and STRADα/β proteins; however, this complex has yet to be characterized in skeletal muscle. In this report, we demonstrate the expression of the LKB1-MO25-STRAD AMPKK complex in adult skeletal muscle, confirm the absence of mRNA splice variants, and report the relative mRNA expression levels of these complex-forming proteins. To facilitate this characterization we used control (ctrl) and muscle-specific LKB1 knockout (LKB1-/-) mice. LKB1 detection in untreated ctrl and LKB1-/- muscle lysates revealed two protein bands at approximately 50 and 60 kDa; although, …
The Expression Of Matrix Metalloproteinase-9 And -2 In Olfactory Injury And Recovery, Stephen Bakos
The Expression Of Matrix Metalloproteinase-9 And -2 In Olfactory Injury And Recovery, Stephen Bakos
Theses and Dissertations
The olfactory system has the remarkable capacity for neurogenesis following injury. However, the molecular mechanisms important for reinnervation of the olfactory bulb (OB) remain unknown. The matrix metalloproteinases (MMPs) are important components in many central nervous system (CNS) injury paradigms, yet remain unexplored in olfactory injury and recovery. To address the role of MMPs, the temporal expressions of MMP-9 and MMP-2 were examined in 3 olfactory injury models: nerve transection (NTx), methyl bromide gas (MeBr) exposure, and nerve transection with Teflon barrier (NTx-TB). Each injury model represents a different degree of olfactory injury and neuronal recovery. In NTx, sensory axons …
Role Of Ampk In The Upregulation Of Steroidogenic Acute Regulatory Protein In The Zona Fasciculata Of The Adrenal Cortex, Adam Wesley Dayton
Role Of Ampk In The Upregulation Of Steroidogenic Acute Regulatory Protein In The Zona Fasciculata Of The Adrenal Cortex, Adam Wesley Dayton
Theses and Dissertations
Cortisol is a glucocorticoid produced by the zona fasciculata (ZF) of the adrenal cortex. Traditionally, cortisol production and release was seen as being regulated strictly by adrenocorticotropic hormone (ACTH). While this is true of baseline cortisol levels and in response to acute mental stress, the picture is somewhat more complicated in other situations.Interleukin-6 (IL-6) contributes to the maintenance of cortisol levels in situations of prolonged immune or inflammatory stress. AMP activated protein kinase (AMPK) was investigated as a possible mediator of the action of IL-6 or as an independent actor in raising cortisol levels in response to hypoxemic or hypoglycemic …
In Vivo Measurement Of Rat Skeletal Muscle Oxygen Consumption Following Brief Periods Of Ischemia With Reperfusion As Assessed By Phosphorescence Quenching Microscopy, William Nugent
Theses and Dissertations
Brief periods of skeletal muscle ischemia (ischemic pre-conditioning) alter cellular metabolism in a way that confers protection over subsequent ischemic episodes. The mechanisms behind this effect have been studied indirectly through assays for the byproducts of ATP synthesis and in vitro studies of cellular signaling cascades and ROS generation. There have been no direct, in vivo assessments of the changes in respiration during reperfusion. We employed phosphorescence quenching microscopy in conjunction with a flow-arrest technique to assess the influences of external, pressure-induced 1- to 10-min focal ischemia on interstitial oxygenation (PISFO2) and the consumption of oxygen (VO2) in spinotrapezius muscles …
Discovery Of A Novel Ccr5 Antagonist As An Effective Therapeutic Agent For Prostate Cancer, Tasrif Ahmed
Discovery Of A Novel Ccr5 Antagonist As An Effective Therapeutic Agent For Prostate Cancer, Tasrif Ahmed
Theses and Dissertations
Previously, the CCR5 receptor was found to be a good target for treating prostate cancer (PCa). Dr. Yan Zhang’s laboratory designed several CCR5 antagonists, which were screened for their inhibitory effect on the growth and invasion of the M12, DU145 and PC-3 PCa cell lines. Primary in vitro screening showed one compound (Drug 17) significantly inhibited the proliferation of PCa cells at 1μM concentration, with a half-maximal inhibitory concentration of 237.68 nM. Further in vitro assays including a proliferation, cytotoxicity and invasion assay confirmed the inhibitory effect of drug 17. The physiological effect of drug 17 was tested by the …
Characterization And Crystallization Of The Mycobacterium Tuberculosis Trmd, Zohal Hamidi
Characterization And Crystallization Of The Mycobacterium Tuberculosis Trmd, Zohal Hamidi
Theses and Dissertations
One third of the world’s population is affected by Tuberculosis (TB), a disease caused by infection with Mycobacterium tuberculosis (MtB). The emergence of multidrug-resistant MtB makes this disease a major public health concern. New agents are needed to treat TB infections in a manner that circumvents existing pathways of resistance. One strategy is to target the organism at the translational level by inhibiting vital modifications of RNA. One gene responsible for these modifications is the tRNA (guanosine-1)-methyltransferase, trmD, which has been shown to be essential in several bacteria. The eukaryotic and bacterial m1G methyltransferases are structurally dissimilar, making this enzyme …
The Effect Of Cobalt Protoporphyrin And Cobalt Chloride On Heme Oxygenase Expression And Protection From Deoxycholate-Induced Apoptosis, Tina Lawson
Theses and Dissertations
The inner surface of the stomach is lined by a mucous membrane known as the gastric mucosa. The integrity of the gastric mucosa is critical for protecting the stomach from the low pH and proteolytic environment within the lumen. Both clinically and experimentally, exposure of gastric mucosal cells to bile salts is known to cause injury. Bile salts present in duodenogastric reflux are thought to play a significant role in gastric ulcer formation and alkaline gastritis. In vitro, studies using physiologic concentrations of the secondary bile salt, deoxycholic acid, indicate that bile salts can induce apoptosis in cultured human gastric …
Molecular Mechanisms Involved Involved In The Interaction Effects Of Hcv And Ethanol On Liver Cirrhosis, Ryan Fassnacht
Molecular Mechanisms Involved Involved In The Interaction Effects Of Hcv And Ethanol On Liver Cirrhosis, Ryan Fassnacht
Theses and Dissertations
The leading causes of liver disease are Hepatitis C virus infection and chronic alcohol abuse. Alcohol accelerates liver disease in HCV but the mechanisms are poorly understood. The identification of molecular gene expression profiles on human liver tissue was performed using microarrays. Samples were obtained from alcoholic-cirrhotic, HCV-cirrhotic, HCV/alcohol-cirrhotic and control non-cirrhotic liver tissue. Probe set expression summaries were calculated using RMA. Probe set level linear models were fit where probe set expression was modeled by HCV status, alcohol status, and the interaction between HCV and Alcohol. HCV cirrhosis was associated with up-regulation of genes related to viral and immune …
Anti-Depressive And Anti-Obesity Changes Following Either Dietary Isoflavone Treatment Or Injection Treatment With The Isoflavonoid Equol: Positive Response Dependent On Animal Age And Ovarian Status In Female Long Evans Rats, Crystal Blake
Theses and Dissertations
Two conditions associated with ovarian depletion are increased potential for depressive episodes and increased abdominal weight gain. In five different experiments we examined the effect of soy-containing diets or equol injections on depression, serotonin levels, weight gain (BW) and white adipose tissue (WAT) deposition of female Long-Evans rats in various stages of life. Rats were intact, ovariectomized or experienced natural ovarian failure (NOF). While this paper will present each experiment, only experiment 5 is outlined here due to space limitations. From conception the rats were exposed to either a soy-rich (Phyto-600) or low-soy diet (Phyto-low). Animals experienced NOF at approximately …
Chronic Amp-Activated Protein Kinase Activation And A High-Fat Diet Have An Additive Effect On Mitochondria In Rat Skeletal Muscle, Natasha Fillmore
Chronic Amp-Activated Protein Kinase Activation And A High-Fat Diet Have An Additive Effect On Mitochondria In Rat Skeletal Muscle, Natasha Fillmore
Theses and Dissertations
Factors that stimulate mitochondrial biogenesis in skeletal muscle include AMPK, calcium, and circulating FFAs. Chronic treatment with either AICAR, a chemical activator of AMPK, or increasing circulating FFAs with a high fat diet increases mitochondria in rat skeletal muscle. The purpose of this study was to determine whether the combination of chronic chemical activation of AMPK and high fat feeding would have an additive effect on skeletal muscle mitochondria levels. We treated Wistar male rats with a high fat diet (HF), AICAR injections (AICAR), or a high fat diet and AICAR injections (HF+AICAR) for six weeks. At the end of …
Liver Kinase B1/Amp-Activated Protein Kinase Signaling In The Diaphragm, Jacob D. Brown
Liver Kinase B1/Amp-Activated Protein Kinase Signaling In The Diaphragm, Jacob D. Brown
Theses and Dissertations
The Liver Kinase B1 (LKB1)/AMP-Activated Protein Kinase (AMPK) signaling pathway is a major regulator of skeletal muscle metabolic processes. During exercise, LKB1-mediated phosphorylation of AMPK leads to its activation, promoting mitochondrial biogenesis and glucose transport, among other effects. The roles of LKB1 and AMPK have not been fully characterized in the diaphragm. Two methods of AMPK activation were used to characterize LKB1/AMPK signaling in diaphragms from muscle-specific LKB1 knockout (KO) and littermate control (C) mice: (1) acute injection of 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) and (2) 5-min direct electrical stimulation (ES) of the diaphragm. Diaphragms were excised 60 minutes post-AICAR injection and …
Role Of Capsule In The Interaction Of Porphyromonas Gingivalis With Host, Amrita Singh
Role Of Capsule In The Interaction Of Porphyromonas Gingivalis With Host, Amrita Singh
Theses and Dissertations
Periodontal disease is a chronic oral disease that is triggered by bacteria. One of these bacteria is Porphyromonas gingivalis. Some strains of P. gingivalis produce capsule, however, so far the role of capsule in the interactions with host cells in P. gingivalis is not well understood. Here, we investigated the contribution of capsule to triggering host response as well as its protective role on bacterial internalization by host cells with subsequent killing. qRT-PCR analysis showed more upregulation of expression of various groups of genes in macrophages challenged with the non-capsulated strain than in those challenged with the capsulated one with …
Signaling Pathways Coupled To Melatonin Receptor Mt1 In Gastric Smooth Muscle, Rashad Ahmed
Signaling Pathways Coupled To Melatonin Receptor Mt1 In Gastric Smooth Muscle, Rashad Ahmed
Theses and Dissertations
The Melatonin, a close derivative of serotonin, is involved in physiological regulation of circadian rhythms. In the gastrointestinal (GI) system, melatonin exhibits endocrine, paracrine and autocrine actions and is implicated in the regulation of GI motility. Generally, melatonin actions oppose the stimulatory actions of serotonin on motility. However, it is not known whether melatonin can also act directly on GI smooth muscle cells. The aim of the present study was to determine the expression of melatonin receptors in smooth muscle and identify their signaling pathways. Muscle cells were isolated from rabbit distal stomach by enzymatic digestion, filtration and centrifugation and …
The Clinical Utility Of Cardiopulmonary Exercise Testing In Patients With Suspected Myocardial Ischemia, Sherry Pinkstaff
The Clinical Utility Of Cardiopulmonary Exercise Testing In Patients With Suspected Myocardial Ischemia, Sherry Pinkstaff
Theses and Dissertations
Heart disease is a major cause of morbidity and mortality in the United States with coronary artery disease (CAD) representing more than half of all cardiovascular events. Stable patients presenting with symptoms suggestive of CAD are likely to undergo either an exercise ECG and/or imaging study as a first line diagnostic assessment. A cardiopulmonary exercise test (CPX) is an ECG stress test plus ventilatory gas analysis. Recently CPX has been used to detect exercise-induced myocardial ischemia suggestive of underlying CAD. Currently there are a number of diagnostic tests available for the identification of CAD with the most widely used being …
Opioid Addiction And Pregnancy: Potential Effects Of Substitution Therapies On Developmental Myelination, Andrew Eschenroeder
Opioid Addiction And Pregnancy: Potential Effects Of Substitution Therapies On Developmental Myelination, Andrew Eschenroeder
Theses and Dissertations
While most cells of the central nervous system (CNS) express opioid receptors, the role of the endogenous opioid system in CNS development remains poorly understood. Identification of opioid functions during brain maturation is particularly crucial in light of the increasing trend in opioid abuse and the use of opioid drugs during pregnancy. New substitution therapies in pregnant opioid addicts include buprenorphine, a mu opioid receptor partial agonist and kappa opioid receptor antagonist. However, while clinical studies demonstrated buprenorphine efficacy in reducing neonatal withdrawal symptoms, there is a lack of information on the potential effects of this drug on brain development. …
Effect Of Hyaluronidase Treatment On The Structural Integrity Of The Endothelial Glycocalyx Layer, Kristin Simmons
Effect Of Hyaluronidase Treatment On The Structural Integrity Of The Endothelial Glycocalyx Layer, Kristin Simmons
Theses and Dissertations
The endothelial glycocalyx plays an important role as part of the permeability barrier between the blood and the interstitium. In this study, we used different sized fluorescently labeled dextran molecules to determine the size of the macromolecular exclusion zone in capillaries. The width of the exclusion zone was calculated as one half the difference between the anatomic luminal diameter, as determined by transillumination, and the width of a fluorescent dextran column. During the first hour after systemic injection of labeled dextrans, neither 70 kDa dextran (Dextran 70) nor 500 kDa dextran (Dextran 500) labeled with the anionic fluorescein isothiocyanate (FITC) …
Role Of Neutrophils In Enhancing Vascular Reactivity To Angiotensin Ii In Preeclampsia, Nikita Mishra
Role Of Neutrophils In Enhancing Vascular Reactivity To Angiotensin Ii In Preeclampsia, Nikita Mishra
Theses and Dissertations
Women with preeclampsia have enhanced vascular reactivity to Angiotensin II (Ang II) and extensive vascular infiltration of neutrophils. The primary mechanism to enhance vessel reactivity is RhoA kinase that phosphorylates MYPT1 to inhibit myosin light chain (MLC) phosphatase. Therefore, MLCs remain phosphorylated and increase sensitivity to calcium. Neutrophils release reactive oxygen species (ROS), which can activate this pathway, so we hypothesized that neutrophils would enhance vessel reactivity to Ang II. Omental vessels from normal pregnant women were used to study vascular reactivity. Ang II dose response (0.001-10µM) was significantly enhanced with perfusion of neutrophils (<2000/mm3, activated with IL-8) or ROS. Addition of superoxide dismutase (SOD)/Catalase to quench ROS or 3µM Y-27632, a specific RhoA kinase inhibitor, blocked enhancement. Vascular smooth muscle expression of pMYPT1 and pMLC in cell culture was significantly increased by neutrophils or ROS. The increase was prevented by Y-27632. RhoA kinase activity assay showed a 3-fold increase in RhoA kinase activity in omental vessels treated with ROS. Similarly, ROS also enhanced vessel reactivity to another vasoconstrictor, norepinephrine, via RhoA kinase. In preeclamptic women, increased neutrophil infiltration is associated with increased vascular expression and production of matrix metalloproteinase-1 (MMP-1). MMP-1 activates protease activated receptor-1 (PAR-1), which could cause endothelial endothelin-1 release, so we considered a novel hypothesis that MMP-1 might cause vasoconstriction and enhance vessel reactivity to Ang II via PAR-1. Omental vessels perfused with activated MMP-1 (0.025-25ng/ml) showed dose-dependent vasoconstriction. Perfusion of activated MMP-1 (2.5ng/ml) significantly enhanced dose response to angiotensin II. MMP-1 mediated vasoconstriction and enhanced vessel reactivity to Ang II was abolished by co-perfusion of 10µM SCH-79797, a specific PAR-1 blocker, and by 5µM BQ-123, a specific endothelin-1 type A receptor blocker. These data are the first to show that activated neutrophils enhance vascular reactivity to Ang II via ROS and the RhoA kinase pathway. They are also the first to show that MMP-1 induces vasoconstriction and enhances vessel reactivity to Ang II. Thus, vascular neutrophil infiltration leading to ROS and MMP-1 generation could be an important mechanism for hypertension in preeclampsia.
The Development Of A Novel Multi-Dimensional Product For Wound Healing Applications, Necrisha Roach
The Development Of A Novel Multi-Dimensional Product For Wound Healing Applications, Necrisha Roach
Theses and Dissertations
A characteristic feature of chronic wounds is a prolonged inflammatory response as well as susceptibility to infection. Studies have shown that during the inflammatory response, there is a significant increase in the levels of neutrophil-derived enzymes. The purpose of this work was to determine whether the anionic macromolecule polystyrene sulfonate (PSS) and five of its salt forms, namely PSS-calcium, PSS-chlorhexidine, PSS-doxycycline, PSS-glutathione and PSS-silver are able to inhibit the activity of three of the enzymes whose levels are elevated in chronic wounds: elastase, cathepsin G and myeloperoxidase. In addition to the enzyme inhibition study, the various formulations’ antimicrobial properties were …
Scribble: A Potential Dual Kinase Inhibitor, Steven Christofakis
Scribble: A Potential Dual Kinase Inhibitor, Steven Christofakis
Theses and Dissertations
Extracellular signal-regulated kinases (ERKs) modulate cellular activities in response to extracellular stimuli and play important biological roles. Thus, perturbed kinase pathways induce pathological conditions, such as tumor development. Rit, a novel member of the Ras family GTPases, activase ERK6, and its over-expression confers tumorigenicity. We hypothesized the presence of scaffolding molecules specific to ERK6, similar to other known MAP kinases. We performed yeast two-hybrid assays using ERK6 as bait, and Scribble was identified as a binding partner. Scribble contains 16 LRR domains and four PDZ domains. We performed immunoprecipitation (IP) assays and discovered ERK2 as another binding partner. Surprisingly, no …
Neutrophil Products Control The Expression Of Progesterone Receptors And Matrix Metalloproteinase-1 In The Decidual And Myometrium And Are Possible Regulators Of Premature Labor, Anna Solotskaya
Theses and Dissertations
Neutrophils infiltrate myometrium and decidual tissue prior to parturition. Activated neutrophils release reactive oxygen species (ROS) and tumor necrosis factor α (TNFα), which might increase expression of pro-labor genes such as matrix metalloproteinase-1 (MMP-1), progesterone receptor (PR) A/B ratio, and cause demethylation of DNA. These changes might cause labor. Decidual tissue was obtained from consented, healthy women at term (37+ weeks of gestation) not in labor (no contractions, without cervical effacement), term labor and preterm labor (under 37 weeks of pregnancy). Decidual and myometrial cells in culture were treated with (1) ROS, (2) TNFα, or (3) 5-aza-2’-deoxycytidine. Total RNA was …
Small Molecule Inhibitors Of Mapk And Pi3k Pathways Enhance Mda-7 Lethality In Renal Cell Carcinoma, Patrick Eulitt
Small Molecule Inhibitors Of Mapk And Pi3k Pathways Enhance Mda-7 Lethality In Renal Cell Carcinoma, Patrick Eulitt
Theses and Dissertations
Renal cell carcinoma accounted for an estimated 57,760 new cases and estimated 12,980 deaths in the United States in 2009. Current treatment options for systemic renal cell carcinoma yield markedly low response percentages; however, recent cytokine therapy experiments have produced promising results. A novel adenovirus, Ad.5/3-mda-7, has been synthesized to efficiently infect renal cancer cells with the mda-7 gene. This gene encodes for the cytokine MDA-7/IL-24 that has the ability to specifically target transformed cells. Assays performed with this adenovirus resulted in an increased percentage of cell death in renal cancer cells when compared to infection with Ad.5/3-cmv empty vector. …
P53 Mediated Cell Motility In H1299 Lung Cancer Cells, Mi-Yon Choi
P53 Mediated Cell Motility In H1299 Lung Cancer Cells, Mi-Yon Choi
Theses and Dissertations
Studies have shown that gain-of- function mutant p53, AKT, and NFκB promote invasion and metastasis in tumor cells. Signals transduced by AKT and p53 are integrated via negative feedback between the two pathways. Tumor derived p53 was also indicated to induce NFκB gene expression. Due to the close relationship between p53/AKT and p53/NFκB, we hypothesized that AKT and NFκB can enhance motility in cells expressing mutant p53. Effects on cell motility were determined by scratch assays. CXCL5- chemokine is also known to induce cell motility. We hypothesized that enhanced cell motility by AKT and NFκB is mediated, in part, by …
Interplay Between Keratin And Vimentin Expression In Oral Cancer, Mary Catherine Mcginn
Interplay Between Keratin And Vimentin Expression In Oral Cancer, Mary Catherine Mcginn
Theses and Dissertations
Previous research in our laboratory found that inhibiting expression of vimentin, a marker of epithelial-to mesenchymal transition, inhibited cell growth and motility in vitro and in vivo. Tumors derived from vimentin knockdown cells showed features of epithelial redifferentiation and increased expression of differentiation-specific keratins. It is unknown what causes re-expression of keratins when vimentin is inhibited. Although, canonical Wnt signaling may activate NF-κB and repress of keratin and/or induce vimentin expression through β-catenin. We hypothesize that downregulation of differentiation-specific keratins contributes to tumor progression, mediated directly or indirectly by expression of vimentin. Vimentin-negative HN4 cells were transfected with plasmids encoding …
Periodontal Bacterial-Dna Initiated Immuno-Inflammatory Responses In Human Osteoblastic Cells, Chebel Najib Bou
Periodontal Bacterial-Dna Initiated Immuno-Inflammatory Responses In Human Osteoblastic Cells, Chebel Najib Bou
Theses and Dissertations
Periodontitis is a chronic inflammatory disease initiated by gram negative anaerobic bacteria. These bacteria possess pathogen-associated molecular patterns (PAMPs) that interact with various receptors including Toll like receptors (TLRs). Bacterial DNA (bDNA) is one of the PAMPs mainly recognized by TLR9. Interaction of bDNA and its receptors leads to activation of inflammatory signaling pathways potentially resulting in periodontal bone destruction. The aim of this study was to determine the production of IL- 6 and IL-8 in response to periodontal bDNA from human osteoblastic cells (MG-63). MG- 63 cells were stimulated in duplicate for 20 hours with 100ng/μl of bDNA from …