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Catecholaminergic Control Of Mitogen-Activated Protein Kinase Signaling In Paraventricular Neuroendocrine Neurons In Vivo And In Vitro: A Proposed Role During Glycemic Challenges, Khan, Todd Ponzio, Graciela Sanchez-Watts, B. Stanley, Glenn Hatton, Alan Watts

Departmental Papers (Biology)

No abstract provided.

Let-7 Expression Defines Two Differentiation Stages Of Cancer, Scott Shell, Sun-Mi Park, Amir Reza Radjabi, Robert Schickel, Emily Kistner, David Jewell

Dartmouth Scholarship

The early phases of carcinogenesis resemble embryonic development, often involving the reexpression of embryonic mesenchymal genes. The NCI60 panel of human tumor cell lines can genetically be subdivided into two superclusters (SCs) that correspond to CD95 Type I and II cells. SC1 cells are characterized by a mesenchymal and SC2 cells by an epithelial gene signature, suggesting that SC1 cells represent less differentiated, advanced stages of cancer. miRNAs are small 20- to 22-nucleotide-long noncoding RNAs that inhibit gene expression at the posttranscriptional level. By performing miRNA expression analysis on 10 Type I and 10 Type II cells, we have determined …

Phase I Study Of Medi-534 Of A Live, Attenuated Intranasal Vaccine Against Respiratory Syncytial Virus And Parainfluenza-3 Virus In Seropositive Children, Margarita Gomez, Maurice A. Mufson, Filip Dubovsky, Conor Knightly, Wen Zeng, Genevieve Losonsky

Internal Medicine

A live, attenuated respiratory syncytial virus and parainfluenza virus type 3 vaccine was evaluated in healthy respiratory syncytial virus/parainfluenza virus type 3 seropositive children aged 1 to 9 years. Three cohorts of 40 children were randomized 1:1 to receive 10^4, 10⁵ or 10⁶ median tissue culture infectious dose₅₀ MEDI-534 vaccine of placebo. The vaccine's safety profile was similar to placebo, no viral shedding was detected, and the vaccine was minimally immunogenic.

Diarrea Viral Bovina Y Animales Portadores Del Virus En Hatos Productores De Leche De La Irrigación De Majes, Arequipa, John C. Huaman G., Hermelinda G. Rivera, Mariluz Araínga, Cesar Gavidia Ch., Alberto Manchego S.

Journal Articles: Pharmacology & Experimental Neuroscience

The objective of this study was to determine the prevalence of bovine viral diarrhea virus (BVDV) and persistently infected (PI) animals in dairy herds located in Majes, Arequipa, Peru. In the first phase, 204 bulk tank milk samples from herds were collected and sent to the local commercial processing plant in Majes for detection of antibodies against BVDV by indirect ELISA test. In the second phase, 286 blood samples from 57 were collected strong positive (Optical Density [OD] ≥ 0.900) for the detection of herds antibodies against BVDV and in PI animals by the indirect and capture ELISAs respectively. In …

Keeping Insects At Bay: Beyond Deet, Thomas P. Rooney

Biological Sciences Faculty Publications

No abstract provided.

Comparison Of C. Elegans And C. Briggsae Genome Sequences Reveals Extensive Conservation Of Chromosome Organization And Synteny, Ladeana W. Hiller, Raymond D. Miller, Scott Everet Baird, Asif Chinwalla, Lucinda A. Fulton, Daniel C. Koboldt, Robert H. Waterston

Biological Sciences Faculty Publications

To determine whether the distinctive features of Caenorhabditis elegans chromosomal organization are shared with the C. briggsae genome, we constructed a single nucleotide polymorphism-based genetic map to order and orient the whole genome shotgun assembly along the six C. briggsae chromosomes. Although these species are of the same genus, their most recent common ancestor existed 80-110 million years ago, and thus they are more evolutionarily distant than, for example, human and mouse. We found that, like C. elegans chromosomes, C. briggsae chromosomes exhibit high levels of recombination on the arms along with higher repeat density, a higher fraction of intronic …

Mutations At F637 In The Nmda Receptor Nr2a Subunit M3 Domain Influence Agonist Potency, Ion Channel Gating And Alcohol Action, H. Ren, Ak Salous, J. M. Paul, Rh Lipsky, Robert W. Peoples

Biomedical Sciences Faculty Research and Publications

Background and purpose:

NMDA receptors are important molecular targets of ethanol action in the CNS. Previous studies have identified a site in membrane-associated domain 3 (M3) of the NR1 subunit and two sites in M4 of the NR2A subunit that influence alcohol action; the sites in NR2A M4 also regulate ion channel gating. The purpose of this study was to determine whether mutations at the site in the NR2A subunit corresponding to the NR1 M3 site influence alcohol action and ion channel gating.

Experimental approach:

We investigated the effects of mutations at phenylalanine (F) 637 of the NR2A subunit using …

Energizing Mirna Research: A Review Of The Role Of Mirnas In Lipid Metabolism, With A Prediction That Mir-103/107 Regulates Human Metabolic Pathways, Bernard R. Wilfred, Wang-Xia Wang, Peter T. Nelson

Sanders-Brown Center on Aging Faculty Publications

MicroRNAs (miRNAs) are powerful regulators of gene expression. Although first discovered in worm larvae, miRNAs play fundamental biological roles-including in humans-well beyond development. MiRNAs participate in the regulation of metabolism (including lipid metabolism) for all animal species studied. A review of the fascinating and fast-growing literature on miRNA regulation of metabolism can be parsed into three main categories: (1) adipocyte biochemistry and cell fate determination; (2) regulation of metabolic biochemistry in invertebrates; and (3) regulation of metabolic biochemistry in mammals. Most research into the 'function' of a given miRNA in metabolic pathways has concentrated on a given miRNA acting upon …

Unlv Magazine, Gian Galassi, Vicki Smith, Erin O'Donnell, Lisa Shawcroft, Angela Sablan, Maria Phelan, Beth English, Eric Leake

UNLV Magazine

No abstract provided.

Bifa, A Cyclic-Di-Gmp Phosphodiesterase, Inversely Regulates Biofilm Formation And Swarming Motility By Pseudomonas Aeruginosa Pa14, Sherry L. Kuchma, Kimberly M. Brothers, Judith H. Merritt, Nicole T. Liberati, Frederick M. Ausubel, George A. O'Toole

Dartmouth Scholarship

The intracellular signaling molecule, cyclic-di-GMP (c-di-GMP), has been shown to influence bacterial behaviors, including motility and biofilm formation. We report the identification and characterization of PA4367, a gene involved in regulating surface-associated behaviors in Pseudomonas aeruginosa. The PA4367 gene encodes a protein with an EAL domain, associated with c-di-GMP phosphodiesterase activity, as well as a GGDEF domain, which is associated with a c-di-GMP-synthesizing diguanylate cyclase activity. Deletion of the PA4367 gene results in a severe defect in swarming motility and a hyperbiofilm phenotype; thus, we designate this gene bifA, for biofilm formation. We show that BifA localizes to the inner …

A Feedback-Controlled Dynamic Linear Actuator To Test Foot Withdrawal Thresholds In Rat, Peter Grigg, D R. Robichaud, Geoffrey M. Bove

Biomedical Sciences Faculty Publications

Many experiments performed in somatosensory research rely upon measuring or estimating the force at which an animal lifts its paw in response to a punctate mechanical stimulus. While mechanical stimulators are now available, the vast majority of methods utilize a series of nylon monofilaments (modern renditions of “von Frey hairs” i.e., Wynnes Symmes filaments, Stoelting Corp), each of which produces a characteristic force (Bove et al 2003). The use of these devices is subject to limitations of the accuracy of the devices, as well as experimenter error and bias (Bove 2006; Bove et al 2003; Fruhstorfer et al 2001). These …

Alternative Targets For The Treatment Of Stroke, Craig T. Ajmo Jr.

USF Tampa Graduate Theses and Dissertations

Stroke is cerebrovascular injury that has been reported to be the third leading cause of death and the first leading cause of disability in the world (W. H.O. 2007). Currently, there is only one FDA approved treatment for stroke which is recombinant tissue plasminogen activator. This treatment has a narrow therapeutic window of three hours after ischemic stroke and can adversely cause the production of oxygen free radicals and intracranial hemorrhage. These limitations result in only 2-3% of all stroke victims as being candidates for this therapy as many patients do not arrive at the hospital in time to receive …

Formation Of Native Prions From Minimal Components In Vitro, Nathan R. Deleault, Brent T. Harris, Judy R. Rees, Surachai Supattapone

Dartmouth Scholarship

The conformational change of a host protein, PrPC, into a disease-associated isoform, PrPSc, appears to play a critical role in the pathogenesis of prion diseases such as Creutzfeldt–Jakob disease and scrapie. However, the fundamental mechanism by which infectious prions are produced in neurons remains unknown. To investigate the mechanism of prion formation biochemically, we conducted a series of experiments using the protein misfolding cyclic amplification (PMCA) technique with a preparation containing only native PrPC and copurified lipid molecules. These experiments showed that successful PMCA propagation of PrPScmolecules in a purified system requires accessory polyanion molecules. In addition, we found that …

Alcohol Slows Interhemispheric Transmission, Increases The Flash-Lag Effect, And Prolongs Masking: Evidence For A Slowing Of Neural Processing And Transmission., Sarah A Khan, Brian Timney

Brain and Mind Institute Researchers' Publications

While the alcohol literature is extensive, relatively little addresses the relationship between physiological effects and behavioural changes. Using the visual system as a model, we examined alcohol's influence on neural temporal processing as a potential means for alcohol's effects. We did this by using tasks that provided a measure of processing speed: Poffenberger paradigm, flash-lag, and backward masking. After moderate alcohol, participants showed longer interhemispheric transmission times, larger flash-lags, and prolonged masking. Our data are consistent with the view that alcohol slows neural processing, and provide support for a reduction in processing efficiency underlying alcohol-induced changes in temporal visual processing.

Acute Myocardial Infarction In Systemic Sclerosis Patients: A Case Series, Chris T. Derk, Sergio A. Jimenez

Department of Medicine Faculty Papers

To characterize the clinical manifestations of patients with systemic sclerosis who develop a myocardial infarction (MI), a retrospective review of the medical records of all patients who were admitted to our institution between 1982 and 2002 and had the dual diagnosis of systemic sclerosis and an acute MI was done. From 1,009 systemic sclerosis hospital admissions, 11 (1.09%) were for an acute MI. Three of these patients had normal coronaries, and instead of wall motion abnormalities, left ventricular hypertrophy was the predominant finding of an echocardiography. The odds ratio of finding normal coronaries in systemic sclerosis vs the general population …

Adiponectin Deficiency Increases Leukocyte-Endothelium Interactions Via Upregulation Of Endothelial Cell Adhesion Molecules In Vivo, Raogo Ouedraogo, Yulan Gong, Brett Berzins, Xiandong Wu, Kalyankar Mahadev, Kelly Hough, Lawrence Chan, Barry J. Goldstein, Rosario Scalia

Department of Medicine Faculty Papers

This study reports on what we believe are novel mechanism(s) of the vascular protective action of adiponectin. We used intravital microscopy to measure leukocyte-endothelium interactions in adiponectin-deficient (Ad^–/–) mice and found that adiponectin deficiency was associated with a 2-fold increase in leukocyte rolling and a 5-fold increase in leukocyte adhesion in the microcirculation. Measurement of endothelial NO (eNO) revealed that adiponectin deficiency drastically reduced levels of eNO in the vascular wall. Immunohistochemistry demonstrated increased expression of E-selectin and VCAM-1 in the vascular endothelium of Ad^–/– mice. Systemic administration of the recombinant globular adiponectin domain (gAd) to Ad …

The Flagellum Of Pseudomonas Aeruginosa Is Required For Resistance To Clearance By Surfactant Protein A, Shiping Zhang, Francis X. Mccormack, Roger C. Levesque, George A. O'Toole, Gee W. Lau

Dartmouth Scholarship

Surfactant protein A (SP-A) is an important lung innate immune protein that kills microbial pathogens by opsonization and membrane permeabilization. We investigated the basis of SP-A-mediated pulmonary clearance of Pseudomonas aeruginosa using genetically-engineered SP-A mice and a library of signature-tagged P. aeruginosa mutants. A mutant with an insertion into flgE, the gene that encodes flagellar hook protein, was preferentially cleared by the SP-A(+/+) mice, but survived in the SP-A(-/-) mice. Opsonization by SP-A did not play a role in flgE clearance. However, exposure to SP-A directly permeabilized and killed the flgE mutant, but not the wild-type parental strain. P. aeruginosa …

Trivalent Vaccine Against Botulinum Toxin Serotypes A, B, And E That Can Be Administered By The Mucosal Route, Easwaran Ravichandran, Fetweh H. Al-Saleem, Denise M. Ancharski, Mohammad D. Elias, Ajay K. Singh, Mohammad Shamim, Yujing Gong, Lance L. Simpson

Department of Medicine Faculty Papers

Most reports dealing with vaccines against botulinum toxin have focused on the injection route of administration. This is unfortunate, because a mucosal vaccine is likely to be more efficacious for patients and pose fewer risks to health care workers and to the environment. Therefore, efforts were made to generate a mucosal vaccine that provides protection against the botulinum serotypes that typically cause human illness (serotypes A, B, and E). This work demonstrated that carboxy-terminal peptides derived from each of the three serotypes were able to bind to and penetrate human epithelial barriers in vitro, and there was no cross inhibition …

The Pharmacokinetics Of Taurolidine Metabolites In Healthy Volunteers., Li Gong, Howard E Greenberg, James L Perhach, Scott A Waldman, Walter K. Kraft

Department of Pharmacology and Experimental Therapeutics Faculty Papers

Taurolidine is an experimental antibacterial and antiendotoxic compound whose clinical utility as an antitumor agent is being investigated in human clinical trials. Taurolidine in aqueous solution exists in equilibrium with taurultam. Taurultam is subsequently transformed to taurinamide. The pharmacokinetic profiles of these metabolites are not well established. In this study, 18 healthy volunteers were administered 5.0 g of taurolidine in 250 mL of 5% polyvinylpyrrolidone in water over 2, 1, or 0.5 hours by intravenous infusion in a parallel-group design. All subjects noted discomfort at the infusion site, although there were no serious adverse events. t(max) generally occurred at the …

Electromagnetic Differences In The Brain During Memory Retrieval, Warren Scott Merrifield

Loma Linda University Electronic Theses, Dissertations & Projects

The primary objectives for this experiment were to analyze the neuroanatomical correlates of autobiographical, episodic and semantic memory, use a different paradigm to promote episodic memory retrieval, and employ a different analysis technique to understand retrieval processes. Autobiographical and episodic memories are personal memories from the past. Autobiographical is more general (e.g. a street name of a house growing up) and episodic is more specific to time (e.g. 13th birthday party that took place on a street). For autobiographical and episodic memory retrieval operations, there is no general consensus as to the localization of function, but bilateral activation of the …

Concave Pit-Containing Scaffold Surfaces Improve Stem Cell-Derived Osteoblast Performance And Lead To Significant Bone Tissue Formation., Antonio Graziano, Riccardo D'Aquino, Maria Gabriella Cusella-De Angelis, Gregorio Laino, Adriano Piattelli, Maurizio Pacifici, Alfredo De Rosa, Gianpaolo Papaccio

Department of Orthopaedic Surgery Faculty Papers

BACKGROUND: Scaffold surface features are thought to be important regulators of stem cell performance and endurance in tissue engineering applications, but details about these fundamental aspects of stem cell biology remain largely unclear.

METHODOLOGY AND FINDINGS: In the present study, smooth clinical-grade lactide-coglyolic acid 85:15 (PLGA) scaffolds were carved as membranes and treated with NMP (N-metil-pyrrolidone) to create controlled subtractive pits or microcavities. Scanning electron and confocal microscopy revealed that the NMP-treated membranes contained: (i) large microcavities of 80-120 microm in diameter and 40-100 microm in depth, which we termed primary; and (ii) smaller microcavities of 10-20 microm in diameter …

Inhibition Of Enterococcus Faecium Adherence To Collagen By Antibodies Against High-Affinity Binding Subdomains Of Acm, Sreedhar R Nallapareddy, Jouko Sillanpää, Vannakambadi K Ganesh, Magnus Höök, Barbara E Murray

Faculty, Staff and Student Publications

Strains of Enterococcus faecium express a cell wall-anchored protein, Acm, which mediates adherence to collagen. Here, we (i) identify the minimal and high-affinity binding subsegments of Acm and (ii) show that anti-Acm immunoglobulin Gs (IgGs) purified against these subsegments reduced E. faecium TX2535 strain collagen adherence up to 73 and 50%, respectively, significantly more than the total IgGs against the full-length Acm A domain (28%) (P < 0.0001). Blocking Acm adherence with functional subsegment-specific antibodies raises the possibility of their use as therapeutic or prophylactic agents.

Crystal Structure Of The Vibrio Cholerae Quorum-Sensing Regulatory Protein Hapr, Rukman S. De Silva, Gabriela Kovacikova, Wei Lin, Ronald K. Taylor, Karen Skorupski, F. Jon Kull

Dartmouth Scholarship

Quorum sensing in Vibrio cholerae involves signaling between two-component sensor protein kinases and the response regulator LuxO to control the expression of the master regulator HapR. HapR, in turn, plays a central role in regulating a number of important processes, such as virulence gene expression and biofilm formation. We have determined the crystal structure of HapR to 2.2-Å resolution. Its structure reveals a dimeric, two-domain molecule with an all-helical structure that is strongly conserved with members of the TetR family of transcriptional regulators. The N-terminal DNA-binding domain contains a helix-turn-helix DNA-binding motif and alteration of certain residues in this domain …

P53 Activation By Knockdown Technologies, Mara E. Robu, Jon D. Larson, Aidas Nasevicius, Soraya Beiraghi, Charles Brenner

Dartmouth Scholarship

Morpholino phosphorodiamidate antisense oligonucleotides (MOs) and short interfering RNAs (siRNAs) are commonly used platforms to study gene function by sequence-specific knockdown. Both technologies, however, can elicit undesirable off-target effects. We have used several model genes to study these effects in detail in the zebrafish, Danio rerio. Using the zebrafish embryo as a template, correct and mistargeting effects are readily discernible through direct comparison of MO-injected animals with well-studied mutants. We show here indistinguishable off-targeting effects for both maternal and zygotic mRNAs and for both translational and splice-site targeting MOs. The major off-targeting effect is mediated through p53 activation, as detected …

Essential Elements Of A Defense-Review Of Dna Testing Results, Dan E. Krane

Biological Sciences Faculty Publications

No abstract provided.

Trans-Snare Complex Assembly And Yeast Vacuole Membrane Fusion, Kevin M. Collins, William T. Wickner

Dartmouth Scholarship

cis-SNARE complexes (anchored in one membrane) are disassembled by Sec17p (α-SNAP) and Sec18p (NSF), permitting the unpaired SNAREs to assemble in trans. We now report a direct assay of trans-SNARE complex formation during yeast vacuole docking. SNARE complex assembly and fusion is promoted by high concentrations of the SNARE Vam7p or Nyv1p or by addition of HOPS (homotypic fusion and vacuole protein sorting), a Ypt7p (Rab)-effector complex with a Sec1/Munc18-family subunit. Inhibitors that target Ypt7p, HOPS, or key regulatory lipids prevent trans-SNARE complex assembly and ensuing fusion. Strikingly, the lipid ligand MED (myristoylated alanine-rich C kinase substrate effector domain) or …

Bladder Inflammatory Transcriptome In Response To Tachykinins: Neurokinin 1 Receptor-Dependent Genes And Transcription Regulatory Elements, Ricardo Saban, Cindy Simpson, Rajanikanth Vadigepalli, Sylvie Memet, Igor Dozmorov, Marcia R. Saban

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Background Tachykinins (TK), such as substance P, and their neurokinin receptors which are ubiquitously expressed in the human urinary tract, represent an endogenous system regulating bladder inflammatory, immune responses, and visceral hypersensitivity. Increasing evidence correlates alterations in the TK system with urinary tract diseases such as neurogenic bladders, outflow obstruction, idiopathic detrusor instability, and interstitial cystitis. However, despite promising effects in animal models, there seems to be no published clinical study showing that NK-receptor antagonists are an effective treatment of pain in general or urinary tract disorders, such as detrusor overactivity. In order to search for therapeutic targets that could …

Microrna Genes Are Frequently Located Near Mouse Cancer Susceptibility Loci., Cinzia Sevignani, George A. Calin, Stephanie C. Nnadi, Masayoshi Shimizu, Ramana V. Davuluri, Terry Hyslop, Peter Demant, Carlo M. Croce, Linda D. Siracusa

Department of Microbiology and Immunology Faculty Papers

MicroRNAs (miRNAs) are short 19- to 24-nt RNA molecules that have been shown to regulate the expression of other genes in a variety of eukaryotic systems. Abnormal expression of miRNAs has been observed in several human cancers, and furthermore, germ-line and somatic mutations in human miRNAs were recently identified in patients with chronic lymphocytic leukemia. Thus, human miRNAs can act as tumor suppressor genes or oncogenes, where mutations, deletions, or amplifications can underlie the development of certain types of leukemia. In addition, previous studies have shown that miRNA expression profiles can distinguish among human solid tumors from different organs. Because …

The Pseudomonas Aeruginosa Secreted Protein Pa2934 Decreases Apical Membrane Expression Of The Cystic Fibrosis Transmembrane Conductance Regulator, Daniel P. Maceachran, Siying Ye, Jennifer M. Bomberger, Deborah A. Hogan, Agnieszka Swiatecka-Urban, Bruce Stanton, George A. O'Toole

Dartmouth Scholarship

We previously reported that Pseudomonas aeruginosa PA14 secretes a protein that can reduce the apical membrane expression of the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Here we report that we have used a proteomic approach to identify this secreted protein as PA2934 [corrected], and we have named the gene cif, for CFTR inhibitory factor. We demonstrate that Cif is a secreted protein and is found associated with outer membrane-derived vesicles. Expression of Cif in Escherichia coli and purification of the C-terminal six-His-tagged Cif protein showed that Cif is necessary and sufficient to mediate the reduction in apical membrane expression …

Stress-Induced Changes In Extracellular Dopamine And Serotonin In The Medial Prefrontal Cortex And Dorsal Hippocampus Of Prenatally Malnourished Rats, David J. Mokler, Olga I. Torres, Janina R. Galler, Peter J. Morgane

Biomedical Sciences Faculty Publications

Prenatal protein malnutrition continues to be a significant problem in the world today. Exposure to prenatal protein malnutrition increases the risk of a number of neuropsychiatric disorders in adulthood including depression, schizophrenia and attentional deficit disorder. In the present experiment we have examined the effects of stress on extracellular serotonin (5-HT) and dopamine in the medial prefrontal cortex and dorsal hippocampus of rats exposed in utero to protein malnutrition. The medial prefrontal cortex and dorsal hippocampus were chosen as two limbic forebrain regions involved in learning and memory, attention and the stress response. Extracellular 5-HT and dopamine were determined in …