Digital Commons Network^™

Inhibition Of The Host Translation Shutoff Response By Herpes Simplex Virus 1 Triggers Nuclear Envelope-Derived Autophagy, Kerstin Radtke, Luc English, Christiane Rondeau, David Leib

Dartmouth Scholarship

Macroautophagy is a cellular pathway that degrades intracellular pathogens and contributes to antigen presentation. Herpes simplex virus 1 (HSV-1) infection triggers both macroautophagy and an additional form of autophagy that uses the nuclear envelope as a source of membrane. The present study constitutes the first in-depth analysis of nuclear envelope-derived autophagy (NEDA). We established LC3a as a marker that allowed us to distinguish between NEDA and macroautophagy in both immunofluorescence and flow cytometry. NEDA was observed in many different cell types, indicating that it is a general response to HSV-1 infection. This autophagic pathway is known to depend on the …

Progress Towards Understanding Of Mechanisms Of Action Of Potent Multifunctional Disease Modifying Therapeutics For Parkinson's Disease & Investigating The Methamphetamine-Induced Striatal Microglia Activation., Mrudang M. Shah

Wayne State University Dissertations

PROGRESS TOWARDS UNDERSTANDING OF MECHANISMS OF ACTION OF POTENT MULTIFUNCTIONAL DISEASE MODIFYING

THERAPEUTICS FOR PARKINSON'S DISEASE.

by

MRUDANG MANOJKUMAR SHAH

December 2013

Advisor: Dr. Aloke Dutta

Major: Pharmaceutical Sciences

Degree: Doctor of Philosophy

Our long term goal is to design and develop potent multifunctional disease modifying therapeutics for Parkinson's disease. The objective of my dissertation was to understand the mechanisms of action of some potent small molecules (synthesized in our lab) as a disease modifying Parkinson's disease therapeutic. The objective was achieved by pursuing the following two specific aims:

1. Investigating anti-oxidant and neuroprotective effects of a lead molecule (D-512) …

Hypoxia-Inducing Factors As Master Regulators Of Stemness Properties And Altered Metabolism Of Cancer- And Metastasis-Initiating Cells., Murielle Mimeault, Surinder K. Batra

Journal Articles: Biochemistry & Molecular Biology

Accumulating lines of experimental evidence have revealed that hypoxia-inducible factors, HIF-1α and HIF-2α, are key regulators of the adaptation of cancer- and metastasis-initiating cells and their differentiated progenies to oxygen and nutrient deprivation during cancer progression under normoxic and hypoxic conditions. Particularly, the sustained stimulation of epidermal growth factor receptor (EGFR), insulin-like growth factor-1 receptor (IGF-1R), stem cell factor (SCF) receptor KIT, transforming growth factor-β receptors (TGF-βRs) and Notch and their downstream signalling elements such as phosphatidylinositol 3'-kinase (PI3K)/Akt/molecular target of rapamycin (mTOR) may lead to an enhanced activity of HIFs. Moreover, the up-regulation of HIFs in cancer cells may …

Historical Remedies Versus Current Pathogens : Laboratory Investigation Of The Efficacy Of Traditional Herbal Remedies Against Present-Day Common Causes Of Bacterial Pneumonia, Eibhlin Mccarthy

Theses

Despite the medical advances of recent years, pneumonia is still a common disease in Ireland, accounting for approximately 7% of deaths in this country'. While national databases such as those run by CIDR and SARI ensure that incidences of systemic and notifiable infections such as tuberculosis are recorded, there are currently no available statistics on commonplace respiratory isolates within Ireland. This study analysed the available data on bacterial pathogens isolated from respiratory samples at a 355 bed Irish hospital over a one year period. Seasonal variations and resistance patterns were noted. The most frequently isolated pathogens were Staphylococcus aureus (27%), …

Time-Stepping For Laser Ablation, Harihar Khanal, David Autrique, Vasilios Alexiades

Publications

Nanosecond laser ablation is a popular technique, applied in many areas of science and technology such as medicine, archaeology, chemistry, environmental and materials sciences. We outline a computational model for radiative and collisional processes occurring during ns-laser ablation, and compare the performance of various low and high order time-stepping algorithms.

Identification Of Potential Synthetic Lethal Genes To P53 Using A Computational Biology Approach, Xiaosheng Wang, Richard Simon

Journal Articles: Genetics, Cell Biology & Anatomy

BACKGROUND:

Identification of genes that are synthetic lethal to p53 is an important strategy for anticancer therapy as p53 mutations have been reported to occur in more than half of all human cancer cases. Although genome-wide RNAi screening is an effective approach to finding synthetic lethal genes, it is costly and labor-intensive.

METHODS:

To illustrate this approach, we identified potentially druggable genes synthetically lethal for p53 using three microarray datasets for gene expression profiles of the NCI-60 cancer cell lines, one next-generation sequencing (RNA-Seq) dataset from the Cancer Genome Atlas (TCGA) project, and one gene expression data from the Cancer …

Effects Of Electrical Stimulation And Testosterone In Translational Models Of Peripheral Nerve Injury, Gina Monaco

Dissertations

As functional recovery following peripheral nerve injury is dependent upon successful regeneration and target reconnection, combinatorial treatments that enhance different regeneration events may be required for recovery from severe injuries. The neurotherapeutic effects of electrical stimulation (ES) and gonadal steroids have been demonstrated independently and in combination in extratemporal and intratemporal facial nerve injuries. The goals of the first aim were to develop a reliable intracranial facial nerve crush model and to investigate the therapeutic potential of combining ES with testosterone propionate (TP) in this most proximal injury model. Adult male rats were divided into intracranial sham-operated, intracranial crush, and …

Investigating The Role For Il-21 In Rabies Virus Vaccine-Induced Immunity., Corin L Dorfmeier, Evgeni P Tzvetkov, Anthony Gatt, James P Mcgettigan

Department of Microbiology and Immunology Faculty Papers

Over two-thirds of the world's population lives in regions where rabies is endemic, resulting in over 15 million people receiving multi-dose post-exposure prophylaxis (PEP) and over 55,000 deaths per year globally. A major goal in rabies virus (RABV) research is to develop a single-dose PEP that would simplify vaccination protocols, reduce costs associated with RABV prevention, and save lives. Protection against RABV infections requires virus neutralizing antibodies; however, factors influencing the development of protective RABV-specific B cell responses remain to be elucidated. Here we used a mouse model of IL-21 receptor-deficiency (IL-21R-/-) to characterize the role for IL-21 in RABV …

Comparison Of Heterologous Prime-Boost Strategies Against Human Immunodeficiency Virus Type 1 Gag Using Negative Stranded Rna Viruses., Tessa M Lawrence, Celestine N Wanjalla, Emily A Gomme, Christoph Wirblich, Anthony Gatt, Elena Carnero, Adolfo García-Sastre, Douglas S Lyles, James P Mcgettigan, Matthias J Schnell

Department of Microbiology and Immunology Faculty Papers

This study analyzed a heterologous prime-boost vaccine approach against HIV-1 using three different antigenically unrelated negative-stranded viruses (NSV) expressing HIV-1 Gag as vaccine vectors: rabies virus (RABV), vesicular stomatitis virus (VSV) and Newcastle disease virus (NDV). We hypothesized that this approach would result in more robust cellular immune responses than those achieved with the use of any of the vaccines alone in a homologous prime-boost regimen. To this end, we primed BALB/c mice with each of the NSV-based vectors. Primed mice were rested for thirty-five days after which we administered a second immunization with the same or heterologous NSV-Gag viruses. …

Isolated Laryngeal Leishmaniasis In Immunocompetent Patients: An Underdiagnosed Disease., Salvatore Cocuzza, Alessio Strazzulla, Marilia Rita Pinzone, Stefano Cosentino, Agostino Serra, Rosario Caltabiano, Salvatore Lanzafame, Bruno Cacopardo, Giuseppe Nunnari

Department of Microbiology and Immunology Faculty Papers

We describe a case of isolated primary laryngeal leishmaniasis in an immunocompetent Italian patient with a previous medical history negative for visceral or cutaneous leishmaniasis, presenting with hoarseness. We also summarize the epidemiological, clinical, and diagnostic features and the therapeutic management of other cases of laryngeal leishmaniasis in immunocompetent subjects, described in the literature. Considering the insidious and nonspecific clinical presentation, the increasing number of different forms of mild or underestimated immunosuppressive conditions, and the number of people travelling in endemic zones, along with the ability of Leishmania amastigotes to survive for a long period in the body, we believe …

Two Coiled-Coil Domains Of Chlamydia Trachomatis Inca Affect Membrane Fusion Events During Infection., Erik Ronzone, Fabienne Paumet

Department of Microbiology and Immunology Faculty Papers

Chlamydia trachomatis replicates in a parasitophorous membrane-bound compartment called an inclusion. The inclusions corrupt host vesicle trafficking networks to avoid the degradative endolysosomal pathway but promote fusion with each other in order to sustain higher bacterial loads in a process known as homotypic fusion. The Chlamydia protein IncA (Inclusion protein A) appears to play central roles in both these processes as it participates to homotypic fusion and inhibits endocytic SNARE-mediated membrane fusion. How IncA selectively inhibits or activates membrane fusion remains poorly understood. In this study, we analyzed the spatial and molecular determinants of IncA's fusogenic and inhibitory functions. Using …

The Tip Of The Tail Needle Affects The Rate Of Dna Delivery By Bacteriophage P22., Justin C Leavitt, Lasha Gogokhia, Eddie B Gilcrease, Anshul Bhardwaj, Gino Cingolani, Sherwood R Casjens

Department of Biochemistry and Molecular Biology Faculty Papers

The P22-like bacteriophages have short tails. Their virions bind to their polysaccharide receptors through six trimeric tailspike proteins that surround the tail tip. These short tails also have a trimeric needle protein that extends beyond the tailspikes from the center of the tail tip, in a position that suggests that it should make first contact with the host's outer membrane during the infection process. The base of the needle serves as a plug that keeps the DNA in the virion, but role of the needle during adsorption and DNA injection is not well understood. Among the P22-like phages are needle …

Attachment Ability And Melanoma Inhibitory Activity Mrna Expression Level Changes In Murine B16-F10 Melanoma Cells Post Nanosecond Electric Pulses, Hongxia Jia

Theses and Dissertations in Biomedical Sciences

The effects of high-voltage nanosecond electric pulses (nsEPs) on metastatic melanoma are still unclear. Hence, we applied one, two, three, and four 300 ns 40 kV/cm pulses to murine B16-F10 melanoma cells. Cell attachment ability was determined by comparing the number of floating cells and the percentage of attached cells. Melanoma inhibitory activity (MIA) is a secretory protein that is highly correlated with the malignancy and metastasis of malignant melanomas. We used MIA as our target to evaluate the effect of nsEPs on metastasis. Pulsed (experimental) and unpulsed (control) cells were incubated at 37°C under a 5% CO₂ atmosphere. …

Neisseria Oralis Sp. Nov., Isolated From Healthy Gingival Plaque And Clinical Samples, William J. Wolfgang, Teresa V. Passaretti, Reashma Jose, Jocelyn Cole, An Coorevits, Andrea N. Carpenter, Sherly Jose, Anita Van Landschoot, Jacques Izard, Donna J. Kohlerschmidt, Peter Vandamme, Floyd E. Dewhirst, Mark A. Fisher, Kimberlee A. Musser

Department of Food Science and Technology: Faculty Publications

A polyphasic analysis was undertaken of seven independent isolates of gram-negative cocci collected from pathological clinical samples from New York, Louisiana, Florida, and Illinois and healthy subgingival plaque from a patient in Virginia, USA. The 16S rRNA gene sequence similarity among these isolates was 99.7–100 %, and the closest species with a validly published name was Neisseria lactamica (96.9 % similarity to the type strain). DNA–DNA hybridization confirmed that these isolates are of the same species and are distinct from their nearest phylogenetic neighbor, N. lactamica. Phylogenetic analysis of 16S and 23S rRNA gene sequences indicated that the …

Manipulation Of Cellular Processing Bodies And Their Constituents By Viruses, Asit K. Pattnaik, Phat X. Dinh

School of Veterinary and Biomedical Sciences: Faculty Publications

The processing bodies (PBs) are a form of cytoplasmic aggregates that house the cellular RNA decay machinery as well as many RNA-binding proteins and mRNAs. The PBs are constitutively present in eukaryotic cells and are involved in maintaining cellular homeostasis by regulating RNA metabolism, cell signaling, and survival. Virus infections result in modification of the PBs and their constituents. Many viruses induce compositionally altered PBs, while many others use specific components of the PBs for their replication. PB constituents are also known to restrict virus replication by a variety of mechanisms. Further, continuing studies in this rapidly emerging field of …

Deep Sequencing And Genome-Wide Analysis Reveals The Expansion Of Microrna Genes In The Gall Midge Mayetiola Destructor, Chitvan Khajuria, Christie E. Williams, Mustapha El Bouhssini, R. Jeff Whitworth, Stephen Richards, Jeffrey J. Stuart, Ming-Shun Chen

School of Veterinary and Biomedical Sciences: Faculty Publications

Background: MicroRNAs (miRNAs) are small non-coding RNAs that play critical roles in regulating post transcriptional gene expression. Gall midges encompass a large group of insects that are of economic importance and also possess fascinating biological traits. The gall midge Mayetiola destructor, commonly known as the Hessian fly, is a destructive pest of wheat and model organism for studying gall midge biology and insect – host plant interactions.

Results: In this study, we systematically analyzed miRNAs from the Hessian fly. Deep-sequencing a Hessian fly larval transcriptome led to the identification of 89 miRNA species that are either identical or very …

Impedance Changes In Biceps Brachii Due To Isometric Contractions And Muscle Fatigue Using Electrical Impedance Myography (Eim), Travis Orth

College of Graduate Studies: Theses & Dissertations

Electrical impedance myography (EIM) is a non-invasive technique used to evaluate neuromuscular conditions by using a quantitative parameter called impedance (Z). It relies upon the application and measurement of high-frequency, low-intensity electrical current imbedded over a localized muscle to determine its opposition to current flow. In brevity, impedance determines the resistance of muscle fibers due to changes in muscle composition. The objective of this thesis was to establish a relationship between muscles at rest and during isometric contractions at various force levels(25%, 50%, and 100% of maximum voluntary contraction) of the biceps brachii over a multifrequency spectrum. Impedance measurements due …

An Update On Vitamin D: The Role In Chronic Disease, Clinical Practice Implications, And The Current Status Of Nurse Practitioner Knowledge, Sara E. Robertson

DNP Projects

The discovery that a large portion of the United States population is vitamin D deficient, has led to hundreds of studies in the last decade which aim to discover the role of vitamin D in overall health and to determine what clinical consequences occur in a deficient state. The current clinical knowledge regarding vitamin D deficiency is reviewed, including the definition of vitamin D insufficiency and deficiency and its correlation to chronic disease states; the risk factors and epidemiology of vitamin D deficiency; and appropriate clinical screening and treatment of vitamin D deficiency. A research study completed in 2012 to …

Thc-Mediated Induction Of Δfosb And Its Modulation Of Cb1r Signaling And Adaptation, Lazenka Matthew

Theses and Dissertations

The main psychoactive and therapeutic effects of Δ9-tetrahydrocannabinol (THC) are mediated through cannabinoid type 1 receptors (CB1Rs). The therapeutic uses of THC are mitigated by the development of tolerance to these therapeutic effects, whereas tolerance does not readily develop to some of the side-effects of THC, like motor impairment and reward. The development of tolerance occurs through adaptations at CB1Rs, which include desensitization (G-protein uncoupling) and downregulation (receptor degradation). Brain region-dependent differences in THC-mediated adaptations are proposed to explain the differences in tolerance to various THC-mediated effects. These studies focused on whether ΔFosB, a stable transcription factor, could regulate CB1R …

A Novel Alcohol-Sensitive Position In The M3 Domain Of The Nmda Receptor Glun2b Subunit, Robert W. Peoples

Biomedical Sciences Faculty Research and Publications

No abstract provided.

Sites Of Alcohol Action At The Glun1/Glun2b Nmda Receptor M3-M4 Domain Intersubunit Interfaces, Yuanhao Zhao, M. Wu, Hong Ren, Robert W. Peoples

Biomedical Sciences Faculty Research and Publications

The N-methyl-D-aspartate (NMDA) receptor has been shown to be one of the most important target sites of alcohol in the central nervous system. We and others have identified positions in the third and fourth membrane-associated (M) domains of both GluN1 and GluN2A subunits that influence alcohol sensitivity. In the structural model of the NMDA receptor based upon the related GluA2 receptor, the outward face of the M3 domain of one subunit type is oriented toward the M4 domain of the other subunit type. We recently reported that four pairs of alcohol-sensitive amino acid positions in GluN1/GluN2A NMDA receptors interact at …

Neurocognitive Function Declines Following Migraine Headache In College Students, Marguerite Moore, T. Covassin, K.A. Pfeiffer, R.E. Norris, R.L. Jensen, C.F. Branta

Journal Articles

No abstract provided.

Therapeutic Potential Of Genetically-Engineered Mesenchymal Stem Cells In Multiple Myeloma, Jerome T. Higgs

All ETDs from UAB

ABSTRACT Multiple Myeloma (MM) remains the second most common hematologic malig-nancy occurring in adults, which primarily affects the skeletal system causing severe bone destruction, spinal cord compression and hypercalcemia. Current therapies, which include chemotherapy, radiotherapy, autologouos stem cell transplantation and in some cases surgery, have extended the median survival between 3 and 10 years. However, MM is still incurable and therefore improving current therapies or developing newer ones to reduce the burden of osteolytic damage and extend survival rate will be highly beneficial for patient management. The long-term goal of this study is to develop a novel therapeutic approach to …

Genetic And Biochemical Characterizations Of The Capsular Glycosyltransferases In Streptococcus Pneumoniae Serotype 2, David Boyd Anthony James

All ETDs from UAB

The principle objective of this dissertation is to develop a thorough understanding of the genetics and biochemistry of capsule synthesis. Specifically, it involves the identification and characterization of the capsular glycosyltransferases in Streptococcus pneumoniae serotype 2, with investigations into their regulation. S. pneumoniae (the pneumococcus) is a significant human pathogen that has the ability to persist as a commensal and then transition to a virulent state, the latter of which is a major contributor to global morbidity and mortality. The capsular polysaccharide (CPS) of S. pneumoniae is a protective surface structure that plays an integral role in this disease process. …

Intracellular Quantitation And Pharmacokinetic Modeling Of Nucleoside Analogs In The Female Genital Tract, Amanda Marie James

All ETDs from UAB

Human Immunodeficiency Virus (HIV) is one of the leading infectious diseases in the world. In spite of substantial progress in the advancement of antiretroviral therapy for HIV treatment, new infections outpace the number of infected persons initiating ART. Therefore, the prevention of HIV remains a crucial health issue. Pre-Exposure Prophylaxis, (PrEP), which involves using one or more antiretroviral agents to reduce the risk of HIV infection prior to potential exposure, has shown effectiveness at reducing HIV transmission in study volunteers with different sexual orientation around the world. The clinical pharmacology of antiretroviral drugs, specifically tenofovir (TFV) and emtricitabine (FTC), in …

Marcks Is A Regulator Of Growth, Radiation Sensitivity And Is A Novel Prognostic Factor For Glioblastoma Multiforme, John Jarboe

All ETDs from UAB

Glioblastoma multiforme (GBM) is the most common and deadly primary brain malignancy necessitating improved understanding of GBM biology. In this study, we explore the role of Myristoylated Alanine Rich C-Kinase Substrate (MARCKS) in the context of GBM. We have discovered that the MARCKS protein regulates GBM growth as well as response to radiation therapy through its effects on proliferation, senescence, and DNA repair based on our studies in cell culture and in patient-derived xenograft tumors implanted in mice. Importantly, our analysis of clinical patient data demonstrates that MARCKS is an independent predictor for outcome in GBM patients. Indeed, high MARCKS …

Usp49 And H2b Deubiquitination In Chromatin And Cellular Function, Amanda E. Jones

All ETDs from UAB

Histones are globular proteins which help package and organize DNA in the eukaryotic nucleus. The four core histones, H2A, H2B, H3 and H4, combine with approximately 146 base pairs of DNA to form a nucleosome, the fundamental unit of chromatin. Post-translational modifications to the C-terminal and N-terminal histone tails regulate chromatin structure and function, and play important roles in transcriptional regulation, replication, DNA damage response, and mRNA processing. Histone H2B ubiquitination is an evolutionarily conserved modification with roles in gene expression and DNA damage repair. In addition, H2B ubiquitination may also regulate mRNA splicing. Here, we identify USP49, a cysteine …

The Role Of Histone Deacetylase Inhibitors In Ameliorating Memory Dysfunction Of An Alzheimer's Disease Mouse Model, Mark Kilgore

All ETDs from UAB

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by amyloid beta plaques and neurofibrillary tangles, along with cognitive impairments that progress to dementia and eventually death. The development of drug treatments that rescue memory deficits could be a promising therapy given in the early stages of AD. In the following studies, we tested the potential for systemic treatment with histone deacetylase (HDAC) inhibitors to ameliorate cognitive deficits in a mouse model of AD. Using the APPSwe/PS1dE9 mouse model of AD, we showed that giving HDAC inhibitors systemically for 3 weeks reversed contextual memory deficits, stably maintained memories over a 2-week …

Virus-Induced Inflammation And Central Nervous System Development, Kate Kosmac

All ETDs from UAB

Congenital human cytomegalovirus (HCMV) infection is a complex cause of central nervous system (CNS) disease. Infection with HCMV during pregnancy results in virus transmission across the placenta and infection of the fetus. With a seroconversion rate in pregnant women between 1% and 4%, HCMV is the most frequently transmitted virus from mother to developing child. Each year between 2,000 and 4,000 infants will suffer from long-term neurologic deficits as a result of congenital infection. The well documented CNS sequelae resulting from congenital HCMV infection have described a wide range of damage without a specific set of pathologic markers for disease. …

Cyclic Gmp Phosphodiesterase As A Novel Target For Colorectal Cancer Chemoprevention, Nan Li

All ETDs from UAB

Nonsteroidal anti-inflammatory drugs (NSAIDs) display promising antineoplastic activity for colorectal and other cancers, but toxicity from cyclooxygenase (COX) inhibition limits their long-term use for chemoprevention. However, many investigators have concluded that COX-independent mechanisms are responsible for their antineoplastic activity. We previously reported that cyclic guanosine monophosphate phosphodiesterase (cGMP PDE) is a non-COX target of sulindac sulfide (SS). However, the specific isozyme families involved have not been well studied. Here we demonstrate that SS inhibits the growth of colon tumor cells through a novel mechanism involving inhibition of two cGMP degrading isozymes, PDE5 and PDE10, to activate cGMP-dependent protein kinase (PKG). …