SURFACE PROTEINS OF BACILLUS ANTHRACIS
M Dominique
University Of Chicagocity: Chicago country: United States (us)
Grant 2R01AI069227-06 from National Institute Of Allergy And Infectious Diseases
Abstract: Bacillus anthracis is the causative agent of anthrax, a zoonotic disease transmitted by spores that germinate in the infected host. The resulting vegetative forms of B. anthracis invade and replicate in all host organs, triggering a lethal outcome to infection and subsequently generating spores for dissemination. Formation of a poly-D-3-glutamic acid (PDGA) capsule represents a key virulence strategy of anthrax bacilli. Further, B. anthracis elaborates a protein surface (S)-layer that is composed of twenty-four S-layer proteins, which are required for this microbe´s virulence. The functions of S-layer proteins include the adherence of bacilli to host tissues, the scavenging of heme-iron, and the control of B. anthracis chain length as an escape from phagocytic clearance. S-layer assembly is initiated via precursor transport across the bacterial plasma membrane by a specialized Sec machine. S-layer proteins subsequently associate via their S-layer homology (SLH) domains with pyruvylated / acetylated secondary cell wall polysaccharide (SCWP), a carbohydrate that is linked to peptidoglycan. S-layer proteins likely contribute to capsule assembly. In B. anthracis, linear PDGA strands are linked to peptidoglycan and threaded across the S-layer. Bacillus cereus G9241, which causes anthrax-like respiratory disease, elaborates a hyaluronic acid capsule; its genetic determinants are conserved in B. anthracis. We propose to use molecular genetic approaches and animal models of anthrax pathogenesis to identify key S-layer virulence factors of B. anthracis, to appreciate their molecular contribution to anthrax pathogenesis and to explore their potential for anthrax vaccine development. Our experimental plan entails studies on the chemical composition of the B. anthracis envelope, microscopic analysis of S-layer protein traffic and capsule synthesis, the structural biology of SLH domain association with SCWP, the characterization of the host receptor for B. anthracis and the development of conjugate capsule-S-layer protein vaccines to protect experimental animals against lethal anthrax challenge. Efforts at generating a human anthrax vaccine from purified subunits, for example protective antigen (PA), have failed. Importantly, PA is dispensable for anthrax pathogenesis. An important frontier is the identification of B. anthracis antigen(s) essential for virulence and that, when used as immunogens, elicit anthrax protective immunity. These questions are addressed in our proposal
Keywords: Address; Adherence (attribute); Adhesions; Animal Model; Animals; Anthrax disease; Anthrax Vaccines; Antibiotic Resistance; Antigens; Bacillus (bacterium); Bacillus anthracis; Bacillus cereus; Biochemical; Biological; Biological Process; capsule (pharmacologic); Carbohydrates; Cavia; Cell membrane; Cell Wall; Cells; Chemicals; Complement Factor B; Conjugate Vaccines; Containment; Development; Disease; Elements; Fluorescence Microscopy; frontier; Genes; Genetic; Genetic Determinism; Glutamic Acid; Goals; Heme Iron; Human; Hyaluronic Acid; Immune; Immunity; improved; Infection; inhibitor/antagonist; insight; Invaded; Killings; Laboratories; Length; Link; Lung diseases; Membrane Proteins; Microbe; Microscopic; Molecular; Molecular Genetics; Monitor; Nature; Operon; Organ; Outcome; Pathogenesis; Pathway interactions; Peptide Signal Sequences; Peptidoglycan; Phagocytosis; Physiological; Play; Polysaccharides; prevent; programs; Protein Precursors; Protein S; protein transport; Proteins; Proteoglycan; receptor; Reproduction spores; Research; research study; Role; structural biology; Structure-Activity Relationship; Surface; Therapeutic; Tissues; trafficking; vaccine development; Vaccines; Virulence; Virulence Factors; weapons; Work
Relevance: Efforts at generating a human anthrax vaccine from purified subunits, for example protective antigen (PA), have failed. Importantly, PA is dispensable for anthrax pathogenesis. An important frontier is the identification of B. anthracis antigen(s) essential for virulence and that, when used as immunogens, elicit anthrax protective immunity. These questions are addressed in our proposal
Project start date: 2006-04-01
Project end date: 2016-12-31
Budget start date: 1-JAN-2012
Budget end date: 31-DEC-2012
2R01AI069227-06 (2012): $384624
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to M Dominique
ANIMAL RESEARCH AND IMMUNOLOGY CORE
M Dominique, Phd
University Of Chicagocity: Chicago country: United States (us)
Abstract: The mission of the Great Lakes Regional Center for Biodefense and Emerging Infectious Diseases (GLRCE) Animal Research and Immunology Core (ARIC) is to provide professional research services in support of the development of vaccines, diagnostics and therapeutics against NIAID Category A, B and C agents and to support GLRCE under emergency situations, marshalling wet laboratory infrastructures and biodefense expertise for the RCE network. During the previous funding period, SARC established infrastructures at the University of Chicago and at the Howard Taylor Ricketts Laboratory (located at Argonne National Laboratory), state-of-the-art BSL-2/3 and ABSL-2/3 research space, enabling work and scientific training under standardized, safe laboratory practices on NIAID Category A-C agents. The GLRCE ARIC acquired CDC registration and certification to sustain a flourishing select agent program that already encompasses several different A-C agents Y. pestis (plague), B. anthracis (anthrax), R. typhi (typhus), R. rickettsii (Rocky Mountain Spotted Fever), R. conori (Mediterranean Spotted Fever), and MRSA (drug resistant S. aureus). ARIC uses a web-based portal (www.glrce.org) to advertise and make research services available to users in the research community, which includes virulence studies in animal models, vaccine and immune responses studies, challenge experiments, characterization of immune responses to immunization or infection, preparation of immunological reagents and assays, drug resistance and therapy studies, measurements of microbial replication in vitro and in vivo, and animal pathology studies. For the next funding period, GLRCE ARIC proposes to provide standardized, high quality animal model systems, to develop and sustain aerobiology systems, to develop immunological tests, reagents and correlates of immunity, and to develop and sustain in vitro testing of small molecule inhibitors for antibiotic activity on NIAID category A-C agents
Keywords: Advertising; Animal Experimentation; Animal Model; Animals; Anthrax disease; Antibiotics; Bacillus anthracis; biodefense; Biological Assay; Biological Models; Boutonneuse Fever; Categories; Centers for Disease Control and Prevention (U.S.); Certification; Chicago; Communities; Diagnostic; Drug resistance; Emergency Situation; Emerging Communicable Diseases; Fever; Funding; Great Lakes Region; Immune response; Immunity; Immunization; Immunologic Tests; Immunology; In Vitro; in vitro testing; in vivo; Infection; inhibitor/antagonist; Intervention; Laboratories; Marshal; Measurement; methicillin resistant Staphylococcus aureus (organism); microbial; Mission; Molecular; National Institute of Allergy and Infectious Disease; Online Systems; pathogen; Pathology; Pharmacotherapy; Plague; Preparation; programs; Reagent; Research; Research Infrastructure; research study; Rocky Mountain Spotted Fever; Services; small molecule; Staphylococcus aureus; System; Testing; Therapeutic; Training; Typhus; Universities; vaccine development; Vaccines; Virulence; Work; Yersinia pestis
Budget start date: 1-MAR-2011
Budget end date: 29-FEB-2012
5U54AI057153-08_5872 (2011): $484821
DETECTION AND CONTROL OF EPILEPSY
M Dominique, Professor
Case Western Reserve Universitycity: Cleveland country: United States (us)
Grant 5R01NS060757-04 from National Institute Of Neurological Disorders And Stroke
Abstract: provided by investigator) Epilepsy is one of the most prevalent neurological disorder. Patients with epilepsy experience recurrent seizures that can cause a variety of symptoms ranging from muscle stiffness to loss of consciousness. Partial epilepsy is the most common syndrome in adult epilepsy patients and mesial temporal lobe epilepsy (MTLE) is the most common form of partial epilepsy. Epilepsy is characterized by the abnormal synchronization of large numbers of neurons. Current therapeutic agents cannot control seizures in 25% of all epileptic patients. Although electrical stimulation of the brain has been very effective to suppress some symptoms of Parkinson´s disease, the level of seizure suppression by stimulation has been limited. The reason for this low therapeutic outcome could be attributed to an inadequate target for stimulation, lack of understanding of the mechanisms and non-optimum stimulation parameters. We propose to develop a method to suppress seizures by applying electrical stimulation to the commissural fiber tract that innervates both hippocampi (major epileptogenic foci in MTLE) from a single electrode location. Specifically, we propose to 1) evaluate the effect of low frequency stimulation 2) determine the effect of high frequency stimulation 3) study the mechanisms of the effect of stimulation, 4) analyze the spatial extent of the suppression/excitation effect in an intact hippocampal in-vitro preparation and 5) suppress seizures in a drug-free genetic model of epilepsy in mice. Current therapeutic agents are incapable of controlling seizure activity in 25% of all epileptic patients. The results of this neuro-technology project should provide valuable insights into the mechanisms underlying seizure suppression with electrical stimulation as well as establishing the preliminary design for an effective neural prosthesis for the control of seizures in patients with epilepsy. About 3 million people have been diagnosed with epilepsy and the cost to the society is about $12.5B/year. Yet, only 25% of patients can receive some therapeutic benefits from anti-epileptic medication. We propose to develop an alternative therapeutic modality to surgical resection that relies on electrical stimulation of the brain to control seizures
Keywords: 4-Aminopyridine; Adult; Affect; Animal Model; Animal Testing; Animals; Antiepileptic Agents; Axon; base; Brain; Breeding; cell body (neuron); Cell Nucleus; Cells; Chronic; clinical application; Collaborations; Conflict (Psychology); Contralateral; cost; Deep Brain Stimulation; design; Detection; Development; Diagnosis; Dimensions; Drug resistance; Dyes; Effectiveness; Electric Stimulation; Electrical Stimulation of the Brain; Electrodes; Electrophysiology (science); Engineering; Epilepsy; Excision; experience; Fiber; Free Will; Frequencies (time pattern); Gene Mutation; Genetic; Genetic Models; Goals; Health; Hippocampus (Brain); Human; Image; implantation; In Vitro; in vitro testing; in vivo; in vivo Model; innovation; insight; Laboratories; Literature; Location; Long-Term Depression (Physiology); Measures; Memory; Methods; Microelectrodes; Modality; Modeling; Movement Disorders; Mus; Muscle; Mutation; nervous system disorder; neural prosthesis; Neuraxis; Neurons; neuroprosthesis; neurotechnology; novel; novel strategies; Operative Surgical Procedures; Outcome; Outcomes Research; Parkinson Disease; Partial Epilepsies; Pathway interactions; Patients; Pharmaceutical Preparations; Physiologic pulse; Picrotoxin; Population; postnatal; Potassium Channel Blockers; Preparation; prevent; Prosthesis; Rattus; Recurrence; relating to nervous system; Relative (related person); Research; Research Personnel; research study; Seizures; Side; Slice; Societies; Sodium Channel; Symptoms; synaptic depression; Synaptic Transmission; Syndrome; Technology; Temporal Lobe; Temporal Lobe Epilepsy; Testing; Therapeutic; Therapeutic Agents; Tissues; tool; Transgenic Mice; Unconscious State; voltage
Project start date: 2008-07-01
Project end date: 2012-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-07-279
5R01NS060757-04 (2011): $336569
THERAPIES OF INFECTIONS CAUSED BY GRAM-POSITIVE BACTERIA
M Dominique, Phd
University Of Chicagocity: Chicago country: United States (us)
Abstract: Gram-positive bacterial pathogens inflict an enormous burden of human disease world-wide. This closely related group of microbes includes Bacillus anthracis, the most notorius bioterror agent, as well as Staphylococcus aureus, which, judged by human morbidity, is currently the single most important infectious agent in the United States. Broad dissemination of antibiotic (methicillin) resistant S. aureus (MRSA) strains in American communities implies the return of the pre-antibiotic era unless new therapies can reduce human mortality. B. anthracis has been weaponized and engineered to acquire antibiotic resistance traits that render currently available antibiotics ineffective and human populations defenseless, if they had been exposed to drug-resistant anthrax spores. The GLRCE Research Project 3 proposal addresses the need for new antibiotics by unraveling molecular mechanisms that lead to assembly of siderophores, proteins, capsules or teichoic acids in the cell wall envelope of B. anthracis and S. aureus. Biosynthesis of all four types of compounds is either essential for bacterial growth or absolutely required for the pathogenesis of infection. An interdisciplinary team of researchers at Argonne National Laboratory, the University of Michigan and the University of Chicago applies multiple different technological platforms to focus on these questions bioinformatics, molecular genetics, biochemical purification and assay development, structure determination, organic chemistry and small molecule inhibition, as well as infectious disease modeling. Products of this research are the in depth molecular appreciation of envelope function and pathogenesis in B. anthracis and S. aureus and the identification of small molecule inhibitors that will be tested for their property of antiinfective or antibiotic therapies. The specific aims are 1. Inhibition of capsular biosynthesis in Bacillus anthracis; 2. Inhibition of lipoteichoic acid biosynthesis in Bacillus anthracis and MRSA; 3. Inhibition of iron siderophore biosynthetic pathways in Bacillus anthracis and MRSA; 4. Inhibition of protein assembly pathways in the envelope of Bacillus anthracis and MRSA; 5. Inhibition of siderophore amide hydrolases in B. anthracis and MRSA
Keywords: Address; adenylate; American; Amides; Anabolism; Animal Model; Anthrax disease; Anti-Infective Agents; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; assay development; Bacillus anthracis; Bacillus anthracis spore; Biochemical; biodefense; Bioinformatics; Biological Factors; capsule (pharmacologic); Cell Wall; Chicago; Communities; Development; Drug resistance; Engineering; Enzymes; Gram-Positive Bacteria; Growth; high throughput screening; Homologous Gene; Human; human disease; human morbidity; human mortality; Hydrolase; in vitro Assay; Infection; Infectious Agent; infectious disease model; inhibitor/antagonist; Intervention; Iron; Laboratories; Lead; lipoteichoic acid; Measures; methicillin resistant Staphylococcus aureus (organism); Michigan; Microbe; Molecular; Molecular Genetics; mutant; Organic Chemistry; pathogen; Pathogenesis; Pathway interactions; petrobactin; Physiological; Population; Property; Protein Inhibition; Proteins; Research; Research Personnel; Research Project Grants; Role; Siderophores; small molecule; sortase; Staphylococcus aureus; Structure; System; Teichoic Acids; Testing; Therapeutic; trait; United States; Universities
Relevance: The GLRCE Research Project 3 proposal addresses the need for new antibiotics by unraveling molecular mechanisms that lead to assembly of siderophores, proteins, capsules or teichoic acids in the cell wall envelope of B. anthracis and S. aureus. Biosynthesis of all four types of compounds is either essential for bacterial growth or absolutely required for the pathogenesis of infection
Budget start date: 1-MAR-2011
Budget end date: 29-FEB-2012
5U54AI057153-08_5860 (2011): $630693
NERVE RESHAPING FOR IMPROVED ELECTRODE SELECTIVITY
M Dominique, Professor
Case Western Reserve Universitycity: Cleveland country: United States (us)
Grant 5R01NS032845-14 from National Institute Of Neurological Disorders And Stroke
Abstract: Functional nerve stimulation (FNS) can restore lost function in patients with damaged nervous system as in spinal cord injury or stroke. Electrodes placed directly on the nerves offer many advantages over muscular stimulation, including control of several muscles with one device, low power requirements, and the potential for recording afferent neural signals. Progress in this area, however, has been hampered by the lack of a truly safe and selective electrode for stimulation of, and recording from, peripheral nerves. In the previous grant application cycle, a novel type of electrode has been developed. This electrode either reshapes a nerve into a flat configuration or maintains it in an already flat shape. This Flat Interface Nerve Electrode (FINE) can both selectively record and stimulate nerves and studies have shown that it can be safe. In this competing renewal, we propose to capitalize on the advantage provided by the ability to line-up the fascicles and to get electrical contacts close to every axon within the nerve. This study has been divided into five primary objectives. The first objective is to determine the effect of the rate of reshaping of the nerve on the safety of the electrode. Hybrid electrodes designs that combine silicone with biodegradable polymer can provide a slow rate of reshaping of the electrode and will be implanted in animals. The second aim focuses on the ability of the fine with contacts arrays positioned along the nerve to activate selectively fibers with specific diameters. The capability to restore normal recruitment by selectively activating small fibers and then large fivers is crucial to the success of neuroprosthetics devices. The third aim is to show that the fine electrode can differentiate neural activity from individual fascicles. Blind sources separation algorithms will be used to obtain the fascicular source activity from the voltage obtained at electrode contacts. The fourth aim is to determine the optimal contact configuration for selective stimulation. Computer models and animal´s experiments will be combined to determine the optimum design in both animal and human nerves. Finally, the last aim is to implement an optimal wireless design of the Flat Interface Nerve Electrode (FINE). A 32-channel peripheral nerve electrode will be built by embedding an electronic circuit within a silicone cuff with only two external leads to a battery pack. The control of the stimulation parameters and the charging of the battery will be done by wireless control. The end product of this project will be a peripheral nerve electrode consisting of a wireless electrode for selective neural stimulation and recording and capable of fiber diameter control. This electrode could significantly improve the ability of current neural prosthetic devices to restore neuromuscular function
Keywords: Algorithms; Animal Experiments; Animals; Apoplexy; Applications Grants; Area; Auditory Prosthesis; Axon; biodegradable polymer; biological signal transduction; bioresorbable polymer; blind; brain attack; Caliber; Cats; Cell Communication and Signaling; Cell Signaling; Cerebral Stroke; cerebral vascular accident; Cerebrovascular accident; Cerebrovascular Apoplexy; Cerebrovascular Stroke; Charge; Cochlear Implants; Cochlear Prosthesis; Common Rat Strains; computational modeling; computational models; computational simulation; computer based models; Computer Simulation; computerized modeling; Computerized Models; computerized simulation; Cranial Nerve XII; Data; degradable polymer; design; designing; Detection; Development; Devices; Diameter; Domestic Cats; Electrodes; Electronics; experiment; experimental research; experimental study; Fascicle; Feline Species; Felis catus; Felis domestica; Felis domesticus; Felis sylvestris catus; Fiber; flexibility; functional restoration; Goals; Grant; Grant Proposals; Grants, Applications; Height; Human; Human, General; Hybrids; Hypoglossal Nerve; Hypoglossal nerve structure; Implant; improved; in silico; Individual; interest; Intracellular Communication and Signaling; Investigators; Mammals, Cats; Mammals, Rats; Man (Taxonomy); Man, Modern; Mathematical Model Simulation; Mathematical Models and Simulations; Measures; Models, Computer; Muscle; Muscle Tissue; Myelopathy, Traumatic; Nerve; Nerve Fibers; nerve reshaping; Nervous; Nervous System damage; Nervous System Injuries; Nervous System Trauma; neural; neural prosthesis; neural prosthetic; neural stimulation; Neurological Damage; Neurological Injury; Neurological trauma; neuromuscular function; novel; Patients; Peripheral; Peripheral Nerves; Physiologic; Physiological; Position; Positioning Attribute; programs; Programs (PT); Programs [Publication Type]; Prosthesis; Prosthetic device; Prosthetics; Rat; Rattus; recruit; Recruitment Activity; relating to nervous system; Research; Research Personnel; research study; Researchers; restore function; restore functionality; restore lost function; Safety; Sciatic Nerve; sciatic nerve; Shapes; Si element; Signal Transduction; Signal Transduction Systems; Signaling; Silicon; Silicones; Simulation, Computer based; Source; Spinal cord injured; Spinal cord injuries; Spinal cord injury; Spinal Cord Trauma; Spinal nerve root structure; Spinal Nerve Roots; Spinal Roots; Spinal Trauma; Stretching; Stroke; stroke; Structure; Structure of sciatic nerve; success; System; System, LOINC Axis 4; Technology; Telemetries; Telemetry; Testing; Time; Trauma, Nervous System; Twelfth Cranial Nerve; V (voltage); vagus nerve stimulation; Vascular Accident, Brain; virtual simulation; voltage; wireless; Wireless Technology
Project start date: 1994-06-01
Project end date: 2011-06-30
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2011
5R01NS032845-14 (2009): $402817