Digital Commons Network^™

Pkcα Translocation And Actin Remodeling In Contracting A7r5 Smooth Muscle Cells, Chenwei Li

Theses, Dissertations and Capstones

Recent research indicates that protein kinase C (PKC) plays an important role in smooth muscle contraction. Because PKC activation and specificity of substrate phosphorylation is believed to be associated with the relocalization of the enzyme to specific cell sites, we first investigated the subcellular translocation of PKCα in A7r5 smooth muscle cells by confocal microscopy through use of standard immunohistologic staining and PKCα - enhanced green fluorescent protein (PKCα -EGFP) fusion protein expression. PKCα was diffusely distributed throughout the cytosol in the unstimulated A7r5 cell. Upon stimulation with phorbol 12, 13 dibutyrate (PDBu), PKCα was translocated primarily to either the …

Glutamine Uptake By Neurons: Interaction Of Protons With System A Transporters, Farrukh A. Chaudhry, Dietmar Schmitz, Richard J. Reimer, Peter Larsson, Andrew T. Gray, Roger Nicoll, Michael Kavanaugh, Robert H. Edwards

Biomedical and Pharmaceutical Sciences Faculty Publications

Astrocytes provide the glutamine required by neurons to synthesize glutamate and GABA. However, the mechanisms involved in glutamine transfer from glia to neurons have remained poorly understood. Recent work has implicated the System N transporter SN1 in the efflux of glutamine from astrocytes and the very closely related System A transporters SA1 and SA2 in glutamine uptake by neurons. To understand how these closely related proteins mediate flux in different directions, we have examined their ionic coupling. In contrast to the electroneutral exchange of H⁺ for Na⁺ and neutral amino acid catalyzed by SN1, we now show that …

Actiononaias Ligamentina As A Biomonitor In The Green River: An Unique Approach For Analysis Of Environmental Impacts, Robert Kirkland

Masters Theses & Specialist Projects

Biomonitoring has become an important component in bioassessment programs. It is used to maintain high water quality standards, and determine contaminant levels and biological affects in areas that have been heavily disturbed. The objectives of this research were 1) to improve and apply certain modern biomonitoring techniques and 2) to locate possible contaminant sources affecting the flora and fauna of the Green River and of Mammoth Cave. Actinonaias ligamentina (Lamarck, 1819), a freshwater mussel, was used for interpretation of these impacts as well as refinement of biomonitoring techniques. The mussels were collected in the Lawler Bend region of the Green …

Medical, Social And Legal Implications Of Treating Nausea And Vomiting Of Pregnancy, Robert L. Brent

The Selected Works of Robert Brent

This article will deal with medical, social, and legal implications of treating nausea and vomiting of pregnancy (NVP). Clinical problems occur when the symptoms become exaggerated and result in debilitation, dehydration, and hospitalization. The treatment of NVP in its early stages has the implication that it will prevent the more serious complications, including hospitalization. Therapeutic modalities discussed in this conference that have been used or are being tested are primarily symptomatic treatments (antihistamines, Bendectin (Merrell Dow; Cincinatti, Ohio), phenothiazines, hyponosis, accupressure, relaxation behavioral modification, audiogenic feedback training, newer medications, diet, and nutritional support). Bendectin is probably the most studied medication …

Mechanisms Counteracting Swelling In Brain Cells During Hyponatremia, Herminia Pasantes-Morales, Rodrigo Franco, Benito Ordaz, Lenin D. Ochoa

School of Veterinary and Biomedical Sciences: Faculty Publications

Water gain in the brain consequent to hyponatremia is counteracted by mechanisms that initially include a compensatory displacement of liquid from the interstitial space to cerebrospinal fluid and systemic circulation and subsequently an active reduction in cell water accomplished by extrusion of intracellular osmolytes to reach osmotic equilibrium. Potassium (K⁺), chloride (Cl^–), amino acids, polyalcohols, and methylamines all contribute to volume regulation, with a major contribution of ions at the early phase and of organic osmolytes at the late phase of the regulatory process. Experimental models in vitro show that osmolyte fluxes occur via leak pathways …

Does Malnutrition During The Lactation Period Affect The Growth Of Bones And Teeth? -A Morphometric Study In Rats, Taner Zi̇ylan, Khalil Awadh Murshed, Aynur Emi̇ne Çi̇çekci̇başi

Turkish Journal of Medical Sciences

To evaluate the effect of protein-energy malnutrition (PEM) on the growth of the mandible and incisor teeth as well as to know if the malnutrition produces modifications in the time of the eruption of the teeth in malnourished rats during the lactation period, animals were divided into experimental and control groups. Experimental pups fed by one mother's milk were doubled in their number to be twice to those of the control pups fed by one mother's milk (Park method). The animals were sacrificed at 20 and 40 days of age, and the body and total lengths of the mandible, and …

Effect Of Low Dietary Zinc Intake And Experimental Diabetes On The Zinc And Carbohydrate Metabolism In Rats, Zine Kechrid, Nazan Demi̇r, Cherif Abdennour, Noureddine Bouzerna

Turkish Journal of Medical Sciences

To investigate the effect of low dietary zinc intake and experimental diabetes (IDDM) on the zinc and carbohydrate metabolism, 8-week-old male wealing normal albino (Wistar) rats were fed diets containing either adequate (54mg/kg) or low zinc (1mg/kg) quantities for one week. Ten rats from each group (n=20) were then intraperitoneally injected with alloxan to induce diabetes. The rats were sacrificed after a further three weeks. Body weight gain and food intake were recorded regularly. On day 28, after an overnight fast, the animals were sacrificed and blood glucose, serum insulin, serum cholesterol concentrations, liver glycogen contents, and femur and pancreatic …

Positive And Negative Roles Of Cd72 In B Cell Function., Hsin-Jung Wu, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Regulation of B cell activation depends on integration of signals transmitted by the B cell receptor (BCR) and a variety of co-receptors. CD72 is a B cell co-receptor that is expressed in all stages of B cell development except plasma cells. Ligation of CD72 enhances B cell growth and differentiation. Recently, the class IV semaphoring, CD100, has been identified as the natural ligand for CD72. Cytoplasmic domain of CD72 has been shown to be associated with SHP-1 leading to the proposal that the positive effects of CD72 on B cell response may result from sequestration of negative signals from BCR. …

Protective Effect Of Nitric Oxide In Lung Injury Associated With Cerulein-Induced Acute Pancreatitis, Abdulkadi̇r Bedi̇rli̇, Erdoğan M. Sözüer, Recep Saraymen, Nai̇l Ersöz, Eser Kiliç

Turkish Journal of Medical Sciences

The aim of this study was to investigate the role of NO in the pathogenesis of lung injury associated with pancreatitis, and the relation between malondialdehyde (MDA) and myeloperoxidase (MPO). Forty Wistar male rats were divided into control or pancreatitis groups, and treated with saline, L-arginine (100 mg/kg i.v.), NO donor, or N-nitro-L-arginine methyl ester (L-NAME) (10 mg/kg i.v.), an inhibitor of NO synthase. Pancreatitis was induced in rats intravenously via the administration of cerulein (10 µg/kg/h) for 4 h). Eight hours after the induction of acute pancreatitis (AP), amylase and lipase in plasma significantly increased in all animals in …

The Effects Of Pentoxifylline In Experimental Nerve Injury, Süleyman Baykal, Cavi̇t Boz, Ertuğrul Çakir, Şükrücan H. Baytan, Murat Karakuş, Kayhan Kuzeyli̇

Turkish Journal of Medical Sciences

Several studies have suggested that pentoxifylline, a theophylline-related drug that decreases erytrhocyte stiffness, may be useful in treatment of ischaemia reperfusion injuries of brain, liver and small intestine in animal experiments. Fourteen Wistar-Albino rats weighing between 180-250 g were used for the experiment and divided equally to two groups. Before the injury, electroneuromyographic (ENMG) data were recorded from right and left sciatic nerves. Right sciatic nerves of the seven rats of group 1 were crushed with a clip. Same lesions were performed on seven rats of group 2 and also pentoxifylline (50mg/kg/day) was applied intraperitoneally once in a day for …

Role Of Bcl-2 Family Members In Caspase-Independent Apoptosis Due To Chlamydia Infection, Jean-Luc Perfettini, John C. Reed, Nicole Israel, Jean-Claude Martinou, Alice Dautry-Varsat, David M. Ojcius

All Dugoni School of Dentistry Faculty Articles

Infection with an obligate intracellular bacterium, the Chlamydia trachomatis lymphogranuloma venereum (LGV/L2) strain or the guinea pig inclusion conjunctivitis serovar of Chlamydia psittaci, leads to apoptosis of host cells. The apoptosis is not affected by a broad-spectrum caspase inhibitor, and caspase-3 is not activated in infected cells, suggesting that apoptosis mediated by these two strains of Chlamydia is independent of known caspases. Overexpression of the proapoptotic Bcl-2 family member, Bax, was previously shown to induce caspase-independent apoptosis, and we find that Bax is activated and translocates from the cytosol to the mitochondria in C. psittaci-infected cells. C. psittaci-induced apoptosis is …

Methods And Compositions For Protection Against Bovine Viral Diseases, Subramaniam Srikumaran

School of Veterinary and Biomedical Sciences: Faculty Publications

The present invention relates to methods and compositions for eliciting an immune response against bovine viral epitopes. The methods comprise combining at least one heat shock protein with at least one bovine viral epitope to form a purified epitope/heat shock protein complex and administration of an immune system stimulating amount of the purified epitope/heat shock protein complex. The compositions comprise, a purified epitope/heat shock protein complex comprising at least one bovine viral epitope complexed with at least one heat shock protein, and a pharmaceutically acceptable carrier, diluent or excipient.

Bile Duct Response To Repeated Cerulein Induced Pancreatitis In The Rat, Ali̇ Menteş, Hasan Aydede, Ümi̇t Bayol, Funda Yilmaz

Turkish Journal of Medical Sciences

The typical morphology of common bile duct strictures in chronic pancreatitis suggests a pathogenesis not of encasement, but of mural changes in the common bile duct. Cerulein, 20 µg/kg was administered to male Wistar rats in four subcutaneous doses, hourly for 3 h, and was recorded as one application. Group A served as controls and received subcutaneous saline. Group B1 received one application of Cerulein and was sacrificed after 24 h. Group B2 was similar to B1, but was sacrificed two weeks later. Group B3 was given two applications of Cerulein with an interval of two weeks and was sacrificed …

Staphylococcus Aureus Agr And Sara Functions Are Required For Invasive Infection But Not Inflammatory Responses In The Lung, Geoffrey Heyer, Shahryar Saba, Robert Adamo, William Rush, Grace Soong, Ambrose Cheung, Alice Prince

Dartmouth Scholarship

Staphylococcus aureus strains lacking agr- and sarA-dependent gene products or specific MSCRAMM (microbial surface components recognizing adhesive matrix molecules) adhesins were compared for the ability to activate inflammatory responses in the lung. The mutants were evaluated for virulence in a mouse model of pneumonia and by quantifying their ability to stimulate interleukin-8 (IL-8) and granulocyte-macrophage colony-stimulating factor (GM-CSF) expression in respiratory epithelial cells. In a neonatal mouse, only strains with intact agr and sarA loci were consistently associated with invasive, fatal pulmonary infection (P < 0.001) and sarA was specifically required to cause bacteremia (P < 0.001). The agr and/or sarA mutants were, nonetheless, fully capable of producing pneumonia and were as proficient as the wild-type strain in stimulating epithelial IL-8 expression, a polymorphonuclear leukocyte chemokine, in airway cells. In contrast, agr and especially sarA mutants induced less epithelial GM-CSF expression, and MSCRAMM mutants lacking fibronectin binding proteins or clumping factor A, a ligand for fibrinogen, were unable to stimulate epithelial GM-CSF production. The ability to induce IL-8 expression was independent of the adherence properties of intact bacteria, indicating that shed and/or secreted bacterial components activate epithelial responses. While conserved staphylococcal components such as peptidoglycan are sufficient to evoke inflammation and cause pneumonia, the agr and sarA loci of S. aureus are critical for the coordination of invasive infection of the lungs.

Transcriptional Interference By Independently Regulated Genes Occurs In Any Relative Arrangement Of The Genes And Is Influenced By Chromosomal Integration Position, Susan K. Eszterhas, Eric E. Bouhassira, David I. K. Martin, Steven Fiering

Dartmouth Scholarship

Transcriptional interference is the influence, generally suppressive, of one active transcriptional unit on another unit linked in cis. Its wide occurrence in experimental systems suggests that it may also influence transcription in many loci, but little is known about its precise nature or underlying mechanisms. Here we report a study of the interaction of two nearly identical transcription units juxtaposed in various arrangements. Each reporter gene in the constructs has its own promoter and enhancer and a strong polyadenylation signal. We used recombinase-mediated cassette exchange (RMCE) to insert the constructs into previously tagged genomic sites in cultured cells. This …

Analysis Of Mitotic Microtubule-Associated Proteins Using Mass Spectrometry Identifies Astrin, A Spindle-Associated Protein, Gary J. Mack, Duane A. Compton

Dartmouth Scholarship

We purified microtubules from a mammalian mitotic extract and obtained an amino acid sequence from each microtubule-associated protein by using mass spectrometry. Most of these proteins are known spindle-associated components with essential functional roles in spindle organization. We generated antibodies against a protein identified in this collection and refer to it as astrin because of its association with astral microtubule arrays assembled in vitro. Astrin is approximately 134 kDa, and except for a large predicted coiled-coil domain in its C-terminal region it lacks any known functional motifs. Astrin associates with spindle microtubules as early as prophase where it concentrates at …

Genetic Dissection Of Behavioral And Neurogenomic Responses To Acute Ethanol, Aaron Wolen

Theses and Dissertations

Individual differences in initial sensitivity to ethanol are strongly related to the heritable risk of alcoholism in humans. To elucidate key molecular networks that modulate ethanol sensitivity we performed a systems genetics analysis of ethanol-responsive gene expression in brain regions of the mesocorticolimbic reward circuit (prefrontal cortex, nucleus accumbens and ventral midbrain) across the BXD RI panel, a highly diverse family of isogenic mouse strains before and after treatment with ethanol. Acute ethanol altered the expression of ~2,750 genes in one or more regions and 400 transcripts were jointly modulated in all three. Ethanol-responsive gene networks were extracted with a …

The Effect Of Lead Acetate On The Susceptibility Of C3heb/Fej Mice Infected With The Parasite Trypanosoma Cruzi, Savonna Warren

Masters Theses & Specialist Projects

Lead has been mined by man for centuries with evidence of its use dating back 6000 years. Chronic exposure to lead can result in encephalopathy, anemia, and nephropathy. Today, cases of lead poisoning are uncommon; however, lead continues to be incorporated into the body through subclinical exposures. These low dosages of lead have been shown to have a deleterious effect on the immune system. Experimental animals exposed to low levels of lead are unable to effectively respond to a number of bacterial and viral challenges. The present study focuses on the relationship between lead and parasitic infection. Parasitemia, mortality, serum …

Effects Of Antioxidant Supplementation With Vitamin E And Selenium On Cytokine Production In Mice Infected With Trypanosoma Cruzi, Dong Chen

Masters Theses & Specialist Projects

Dietary supplementation with the antioxidants vitamin E and selenium (Se) has been shown to be beneficial against the intracellular parasite Trypanosoma cruzi during experimental Chagas' disease. Supplementation of mice with vitamin E and selenium at levels well above the recommended daily allowance for these two substances results in significantly decreased parasitemia levels and increased longevity. However, the mechanistic role of the antioxidants in this process is not well understood. It is known that vitamin E and selenium can reduce oxidative stress and improve host immune responses. In addition, the synthesis of many regulatory cytokines is known to be influenced by …

Dextran-Methylprednisolone Succinate As A Prodrug Of Methylprednisolone: Plasma And Tissue Disposition, Xiaoping Zhang, Reza Mehvar

Pharmacy Faculty Articles and Research

Plasma and tissue disposition of a macromolecular prodrug of methylprednisolone (MP), dextran (70 kDa)–methylprednisolone succinate (DMP), was studied in rats. Single 5‐mg/kg doses of DMP or unconjugated MP were administered into the tail veins of different groups of rats (n  = 4/group/time point). Blood (cardiac puncture) and tissues (liver, spleen, kidney, heart, lung, thymus, and brain) were collected at various times after DMP (0–96 h) or MP (0–2 h) injections. Concentrations of DMP and MP in samples were analyzed by size‐exclusion chromatography (SEC) and reversed‐phase high‐performance liquid chromatography (HPLC), respectively. Conjugation of MP with 70‐kDa dextran resulted in 22‐, …

Evaluation Of Cholera Vaccines Formulated With Toxin-Coregulated Pilin Peptide Plus Polymer Adjuvant In Mice, Jia-Yan Wu, William F. Wade, Ronald K. Taylor

Dartmouth Scholarship

Cholera is an acute diarrheal disease that is caused by the gram-negative bacterium Vibrio cholerae. The low efficacy of currently available killed-whole-cell vaccines and the reactinogenicity coupled with potential reversion of live vaccines have thus far precluded widespread vaccination for the control of cholera. Recent studies on the molecular nature of the virulence components that contribute to V. cholerae pathogenesis have provided insights into possible approaches for the development of a defined subunit cholera vaccine. Genetic analysis has demonstrated that the toxin-coregulated pilus (TCP) is the major factor that contributes to colonization of the human intestine by V. cholerae. In …

Immune Response Genes Modulate Serologic Responses To Vibrio Cholerae Tcpa Pilin Peptides, Michael D. Meeks, Terri K. Wade, Ronald K. Taylor, William F. Wade

Dartmouth Scholarship

Cholera is an enteric disease caused by Vibrio cholerae. Toxin-coregulated pilus (TCP), a type 4 pilus expressed by V. cholerae, is a cholera virulence factor that is required for host colonization. The TCP polymer is composed of subunits of TcpA pilin. Antibodies directed against TcpA are protective in animal models of cholera. While natural or recombinant forms of TcpA are difficult to purify to homogeneity, it is anticipated that synthesized TcpA peptides might serve as immunogens in a subunit vaccine. We wanted to assess the potential for effects of the immune response (Ir) gene that could complicate a peptide-based …

Characterization Of Seca-Sod Operon In Borrellia Burgdorferi., Tonya Lynn Nichols

Electronic Theses and Dissertations

Borrelia burgdorferi, the causative agent of Lyme disease, has been characterized as a microaerophilic spirochete. O2 consumption and utilization potentially yield reactive oxygen intermediates, such as superoxide, hydroxyl radicals, and hydrogen peroxide. This study investigated the expression of the sod gene, which encodes the only, identified oxidative defense mechanism in B. burgdorferi. Using primer extension analysis and RT-PCR, it was found that sod and secA are organized as a single transcriptional unit under the control of σ70-like promoter upstream of the secA open reading frame. Generally, gene expression decreases with increased distance from the promoter; however, secA expression was observed …

Behavioral Changes In Adult C57bl/6j Mice Following Prenatal Exposure To Ethanol., Kevin Wade Nunley

Electronic Theses and Dissertations

Fetal Alcohol Syndrome (FAS) labels children with physical, mental and behavioral deficits exposed to alcohol in utero. Current research indicates that timing of alcohol exposure of the embryo/fetus is a critical determinant of the behavioral deficits associated with FAS. This study represents a model for binge drinking, in which C57BL mouse embryos were exposed to alcohol during 2 separate critical periods of brain development, gestational day (GD) 7 or 8. As adults, the offspring were tested to determine if loco-motor activity and emotional reaction to a novel environment had been affected. Significant differences due to treatment and sex were …

Anti-Class Ii Monoclonal Antibody-Targeted Vibrio Cholerae Tcpa Pilin: Modulation Of Serologic Response, Epitope Specificity, And Isotype, Jia-Yan Wu, Ronald K. Taylor, William F. Wade

Dartmouth Scholarship

Toxin-coregulated pilus (TCP) is a colonization factor required for cholera infection. It is not a strong immunogen when delivered in the context of whole cells, yet pilus subunits or TcpA derivative synthetic peptides induce protective responses. We examined the efficacy of immunizing mice with TCP conjugated to anti-class II monoclonal antibodies (MAb) with or without the addition of cholera toxin (CT) or anti-CD40 MAb to determine if the serologic response to TcpA could be manipulated. Anti-class II MAb-targeted TCP influenced the anti-TCP peptide serologic response with respect to titer and isotype. Responses to TcpA peptide 4 were induced with class …

Evaluation Of A Tetracycline-Inducible Promoter In Staphylococcus Aureus In Vitro And In Vivo And Its Application In Demonstrating The Role Of Sigb In Microcolony Formation, B. T. Bateman, N. P. Donegan, T. M. Jarry, M. Palma

Dartmouth Scholarship

An inducible promoter system provides a powerful tool for studying the genetic basis for virulence. A variety of inducible systems have been used in other organisms, including pXyl-xylR-inducible promoter, the pSpac-lacI system, and the arabinose-inducible P_BAD promoter, but each of these systems has limitations in its application to Staphylococcus aureus. In this study, we demonstrated the efficacy of a tetracycline-inducible promoter system in inducing gene expression in S. aureus in vitro and inside epithelial cells as well as in an animal model of infection. Using the xyl/tetO promoter::gfp_uvr fusion carried on a shuttle …

Molecular Studies Involving The Rev Protein Of Caprine Arthritis Encephalitis Virus And Visna Virus., Bridget Michele Graves

Electronic Theses and Dissertations

Caprine Arthirtis Encephalitis Virus (CAEV) and Visna Virus are two viruses of the lentivirus family. They encode three structural genes (gag, pol, and env) and two regulatory genes (rev and tat). The Rev protein regulates Gag, Pol and Env expression by transporting their mRNAs to the cytoplasm by binding to the RRE (Rev Response Element) found on their mRNAs. Previous studies have indicated that Rev may be toxic to transfected cells, overexpression of exogenous RREs or a better binding RRE can inhibit Rev activity and Rev-C (CAEV Rev) can trans-activate RRE-V (Visna Virus …

Effect Of Monounsaturated Fat Rich Almonds On Hemostatic And Inflammatory Factors In Healthy Adults, Kristianne M. Connell

Loma Linda University Electronic Theses, Dissertations & Projects

Background: The frequent consumption of nuts is inversely associated with the incidence of cardiovascular disease. Nuts are known to decrease the risk of cardiovascular disease by reducing serum total and low-density lipoprotein (LDL) cholesterol. Markers of inflammation including C-reactive protein, E-selectin and interleukin-6 have been proposed to be newer risk factors for cardiovascular disease. However, the influence of nuts such as almonds on markers of inflammation and hemostasis that influence cardiovascular disease risk is not currently known.

Objective: The objective of this study was to determine the effect of almond consumption on markers of inflammation and hemostatic factors in healthy …

Proteolytic Release Of Cd44 Intracellular Domain And Its Role In The Cd44 Signaling Pathway., I Okamoto, Y Kawano, D Murakami, T Sasayama, N Araki, T Miki, A J Wong, H Saya

Department of Microbiology and Immunology Faculty Papers

CD44 is a widely distributed cell surface adhesion molecule and is implicated in diverse biological processes. However, the nature of intracellular signaling triggered by CD44 remains to be elucidated. Here, we show that CD44 undergoes sequential proteolytic cleavage in the ectodomain and intracellular domain, resulting in the release of a CD44 intracellular domain (ICD) fragment. Consequently, CD44ICD acts as a signal transduction molecule, where it translocates to the nucleus and activates transcription mediated through the 12-O-tetradecanoylphorbol 13-acetate-responsive element, which is found in numerous genes involved in diverse cellular processes. Expression of an uncleavable CD44 mutant as well as metalloprotease inhibitor …

Protection Against Ischemia And Reperfusion Injury, Peter R. Oeltgen, Paul D. Bishop, Mark S. Kindy, Juan A. Sanchez

Molecular and Cellular Biochemistry Faculty Patents

A compound and method for using the compound to reduce injury associated with ischemia and reperfusion of mammalian organs such as the heart. The compound may be administered as part of a preconditioning strategy which reduces the extent of injury and improves organ function following cessation and restoration of blood flow. The compound may be used in preparation for planned ischemia or in a prophylactic manner in anticipation of further ischemic events.