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Purification And Scale-Up Of A Recombinant Heavy Chain Fragment C Of Botulinum Neurotoxin Serotype E In Pichia Pastoris Gs115, Michael P. Dux, Rick Barent, Jayanta Sinha, Mark Gouthro, Todd Swanson, Ardis Barthuli, Mehmet Inan, John T. Ross, Leonard A. Smith, Theresa J. Smith, Robert Webb, Bonnie Loveless, Ian Henderson, Michael M. Meagher

Michael Meagher Publications

A recombinant C-terminus heavy chain fragment from botulinum neurotoxin serotype E (BoNT/E) is proposed as a vaccine against the serotype E neurotoxin. This fragment, rBoNTE(H_c), was produced intracellular in Pichia pastoris GS115 by a three-step fermentation process, i.e., glycerol batch phase and a glycerol fed-batch phase to achieve high cell densities, followed by a methanol fed-batch induction phase. The rBoNTE(H_c) protein was purified from the soluble fraction of cell lysates using three ion-exchange chromatography steps (SP Sepharose Fast Flow, Q Sepharose Fast Flow, Sp Sepharose High Performance) and polished with a hydrophobic charge induction chromatography step …

Heat Transfer And Chemical Processes In Chemical Vapor Deposition Reactor For Synthesis Of Carbon Nanotubes, Wahed Rezk Wasel

University of Kentucky Doctoral Dissertations

A small-scale model of a CVD reactor was built. Axial and radial of major species concentrations and temperature profiles were obtained with a micro gas chromatograph and a fine thermocouple. Those temperature and species concentrations revealed detailed thermal and chemical structures of the CVD reactor.

The concentrations of argon plus hydrogen, methane, and C₂H_x (C₂H₂ + C₂H₄ + C₂H₆) resulting from xylene decomposition were measured along the CVD at different temperatures. Ferrocene was added to xylene to investigate the effect of a catalyst on composition profiles. The …

Bs News

Building Services Engineering

No abstract provided.

Impulse, Dave Graves, Kyle Johnson, Miranda Reiman, Nicole Schaffer, Greta Stewart, Denise Watt

Impulse (Jerome J. Lohr College of Engineering Publication)

Features:
[Page] 2 South Dakota no longer is merely a launching pad: An industrialization of the economy has made the state a land of opportunity for engineers while still having a small town feel.
[Page] 4 While you were away: Brookings and SDSU aren't the places many alums remember them to be.
[Page] 6 What will lead future growth in the region?: University and community leaders are proposing a 134-acre research park adjacent to the SDSU campus.
[Page] 8 GIS Center of Excellence: Scientists have been hired for the inaugural year of operation for the Geographic Information Science Center of …

Automated Liquid Dispensing Pin For Dna Microarray Applications, Suganya Parthasarathy

Theses

This thesis describes the research and development of a new liquid dispensing/aspiring system that is capable of producing micro sized spots/droplets for molecular biology research and analysis. In particular, the application is focused on DNA microarray fabrication with the goals of smaller spot size, higher yield, more efficient usage of biological materials, and capability to handle high viscosity liquids. The new system is based on active sensing and control and it is part of a fully integrated robotic microarray system for genomic and proteomic applications. The prototype system handles water as well as thick liquids such as 100% glycerol and …

Purification And Scale-Up Of A Recombinant Heavy Chain Fragment C Of Botulinum Neurotoxin Serotype E In Pichia Pastoris Gs115, Michael P. Dux, Rick Barent, Jayanta Sinha, Mark Gouthro, Todd Swanson, Ardis Barthuli, Mehmet İnan, John T. Ross, Leonard A. Smith, Theresa J. Smith, Robert Webb, Bonnie Loveless, Ian Henderson, Michael M. Meagher

Papers in Biochemical Engineering

A recombinant C-terminus heavy chain fragment from Botulinum neurotoxin serotype E (BoNT/E) is proposed as a vaccine against the serotype E neurotoxin. This fragment, rBoNTE(Hc), was produced intracellular in Pichia pastoris GS115 by a three-step fermentation process, i.e., glycerol batch phase and a glycerol fed-batch phase to achieve high cell densities, followed by a methanol fed-batch induction phase. The rBoNTE(Hc) protein was purified from the soluble fraction of cell lysates using three ion-exchange chromatography steps (SP Sepharose Fast Flow, Q Sepharose Fast Flow, Sp Sepharose High Performance) and polished with a hydrophobic charge induction chromatography step (MEP HyperCel). Method development …

Crystallization And Preliminary X-Ray Analysis Of Na-Asp-1, A Multi-Domain Pathogenesis-Related-1 Protein From The Human Hookworm Parasite Necator Americanus, Mehmet İnan, Oluwatoyin A. Asojo, Alex Lucas, Rick Barent, Jicai Huang, Brad Plantz, Amber Swanson, Mark Gouthro, Michael M. Meagher, Peter J. Hotez

Papers in Biochemical Engineering

Human hookworm infection is a major cause of anemia and malnutrition in the developing world. In an effort to control hookworm infection, the Human Hookworm Vaccine Initiative has identified candidate vaccine antigens from the infective larval stage (L₃) of the parasite, including a family of pathogenesis related- 1 (PR-1) proteins known as the ancylostoma-secreted proteins (ASPs). The functions of the ASPs are unknown. In addition, it is unclear why some ASPs have one while others have multiple PR-1 domains. There are no known structures of a multi-domain ASP and in an effort to remedy this situation, recombinant Na …

Conformational Sampling Of The Botulinum Neurotoxin Serotype A Light Chain: Implications For Inhibitor Binding, James C. Burnett, James J. Schmidt, Connor F. Mcgrath, Tam L. Nguyen, Ann R. Hermone, Rekha G. Panchal, Jonathan L. Vennerstrom, Krishna Kodukula, Daniel W. Zaharevitz, Rick Gussio, Sina Bavari

United States Army: Publications

Botulinum neurotoxins (BoNTs) are the most potent of the known biological toxins, and consequently are listed as category A biowarfare agents. Currently, the only treatments against BoNTs include preventative antitoxins and long-term supportive care. Consequently, there is an urgent need for therapeutics to counter these enzymes––post exposure. In a previous study, we identified a number of small, nonpeptidic lead inhibitors of BoNT serotype A light chain (BoNT/A LC) metalloprotease activity, and we identified a common pharmacophore for these molecules. In this study, we have focused on how the dynamic movement of amino acid residues in and surrounding the substrate binding …

Use Of Calcium Hypochlorite As A Sanitizer For Seeds Used For Sprouting: Task #2; Impact: Improved Alfalfa Decontamination Technologies, Emily Damron, Carrie Klein, Melissa Leach, Jordan Mourot, Tom Murphy, Amy Seamans, Ryan Wilson

Inquiry: The University of Arkansas Undergraduate Research Journal

Consumption of raw or lightly cooked alfalfa sprouts has been a concern of the U.S. Food and Drug Administration (FDA) in recent years due to connections between sprouts and foodborne illnesses. Researchers have identified, contaminated seeds as the primary source of alfalfa sprouts contamination. Contamination of alfalfa seeds can originate in the field, harvesting, storing, or sprouting. Two pathogens of particular concern on alfalfa seeds are Escherichia coli 0157:H7 (E. coli) and Salmonella. These pathogens are capable of producing biofilms that provide protection for individual cells and allow for survival in otherwise hostile environments, including some disinfectant washes. Other factors …

Towards An Rts,S-Based, Multi-Stage, Multi-Antigen Vaccine Against Falciparum Malaria: Progress At The Walter Reed Army Institute Of Research, D. Gray Heppner Jr., Kent E. Kester, Christian F. Ockenhouse, Nadia Tornieporth, Opokua Ofori, Jeffrey A. Lyon, V. Ann Stewart, Patrice Dubois, David E. Lanar, Urszula Krzych, Philippe Moris, Evelina Angov, James F. Cummings, Amanda Leach, B. Ted Hall, Sheetij Dutta, Robert Schwenk, Collette Hillier, Arnoldo Barbosa, Lisa A. Ware, Lalitha Nair, Christian A. Darko, Mark R. Withers, Bernhards Ogutu, Mark E. Polhemus, Mark Fukuda, Sathit Pichyangkul, Montip Gettyacamin, Carter Diggs, Lorraine Soisson, Jessica Milman, Marie-Claude Dubois, Nathalie Garcon, Kathryn Tucker, Janet Wittes, Christopher V. Plowe, Mahamadou A. Thera, Ogobara K. Duombo, Maria G. Pau, Jaap Goudsmit, W. Ripley Ballou, Joe Cohen

United States Army: Publications

The goal of the Malaria Vaccine Program at the Walter Reed Army Institute of Research (WRAIR) is to develop a licensed multi-antigen, multi-stage vaccine against Plasmodium falciparum able to prevent all symptomatic manifestations of malaria by preventing parasitemia. A secondary goal is to limit disease in vaccinees that do develop malaria. Malaria prevention will be achieved by inducing humoral and cellular immunity against the pre-erythrocytic circumsporozoite protein (CSP) and the liver stage antigen-1 (LSA-1). The strategy to limit disease will target immune responses against one or more blood stage antigens, merozoite surface protein-1 (MSP-1) and apical merozoite antigen-1 (AMA-1). The …

Fuzzifying [Sic] Markov Decision Process, Ahmed Syed Irshad

LSU Master's Theses

Markov decision processes have become an indispensable tool in applications as diverse as equipment maintenance, manufacturing systems, inventory control, queuing networks and investment analysis. Typically we have a controlled Markov chain on a suitable state space in which transitional probabilities depend on the policy (or decision maker) which comes from a set of possible actions. The main problem of interest would be to find an optimal policy that minimizes the associated cost. Linear Programming has been widely used to find the optimal Markov decision policy. It requires solutions of large systems of simultaneous linear equations. By the fact that the …

Ridding Ships' Ballast Water Of Microorganisms, Fred C. Dobbs, Andrew Rogerson

OES Faculty Publications

( First paragraph) A complex assemblage of microorganisms exists in nearly every aquatic system on earth. In lakes and oceans, every milliliter of water contains about 10² protists (single-celled eukaryotes), 10⁶ bacteria, and 10⁷–10⁹ viruses. Therefore, billions of microorganisms inevitably enter ships’ ballast tanks during normal operations. It has been argued that microorganisms must certainly be frequent invaders of coastal ecosystems, given the high densities of bacteria and viruses in ballast water—10⁸ and 10⁹ organisms per liter, respectively (1)—their potentially high reproductive rates, broad tolerances to physical conditions, and ability to form resting …

Production And Purification Of A Chimeric Monoclonal Antibody Against Botulinum Neurotoxin Serotype A , Mark C. Mowry, Michael Meagher Lead Investigator, Leonard Smith, Anuradha Subramanian

Papers in Biotechnology

Production of recombinant antibodies against botulinum neurotoxin is necessary for the development of a post-exposure treatment. CHO-DG44 cells were transfected with a plasmid encoding the light and heavy chains of a chimeric monoclonal antibody (S25) against botulism neurotoxin serotype A. Stable cell lines were obtained by dilution cloning and clones were shown to produce nearly equivalent levels of light and heavy chain antibody by an enzyme-linked immunosorbent assay (ELISA). In suspension culture, cells produced 35 μg/ml of chimeric antibody after 6 days, corresponding to a specific antibody productivity of 3.1 pg/cell/day. A method for the harvest and recovery of an …

Flight Physician - September, 2004, Civil Aviation Medical Association

Browse all Civil Aviation Medical Association Newsletters

A twenty page newsletter of the Civil Aviation Medical Association. The newsletter provided news about civil aviation medicine and information related to the organization.

Photodynamic Modulation Of Wound Healing And Inhibition Of Tissue Degradation, Michael R Hamblin, Tatiana Demidova, Faten Gad, Touqir Zahra, Tayyaba Hasan

Light Activated Tissue Regeneration and Therapy

No abstract provided.

Particle Size Dependence On Immobilized Molecular Sensors : Assay Of Nucleic Acid Probes Attached To Paramagnetic Particles, Celin Sunny

Theses

Nanotechnology is concerned with materials and systems whose structure and components exhibit novel and significantly improved physical, chemical and biological properties, phenomena and processes due to their nano scale size.

The transition from micro scale to nano scale leads to a number of changes in physical properties, possibly including new physical principles, some of which may be yet to be discovered. One of the major factors in this is the increase in ratio of surface area to volume.

This thesis research uses the increase in ratio of surface area to volume property of going from macro to micro to nano …

Gsbs News, Spring 2003-2004, Graduate School Of Biomedical Sciences, University Of Texas Health Science Center At Houston/M.D. Anderson Cancer Center

GSBS News

No abstract provided.

Nebraska Blueprint - Spring 2004

Nebraska Blueprint (Student Publication)

Table of Contents:

From the Editor
A Well-Earned Yet Charmed Life by Tom Cudd
Combating Tomorrow’s Diseases Today by Kevin Mlnarik
Where We're Headed by Cecelia Orwig
Not Today But Here Tomorrow by Tom Cudd
My So Called Life: 2054 by Cecelia Orwig
What It's All About by Tom Cudd
Why Don't Engineers get involved? by Tom Cudd
ASME Hosts Successful Conference

Carbon Fiber For Air Filtration, Marit Jagtoyen, Francis J. Derbyshire, Rosemary Anne Alexander Derbyshire

Center for Applied Energy Research Faculty Patents

Described are filters containing free-activated carbon fibers or activated carbon fibers in the form of a composite used for filtering a variety of contaminants, including pathogens, from air. These filters can provide effective and efficient removal of contaminants from air, requiring only short contact time between the air being filtered and the filter itself, with only a minimum pressure drop across the filter. The characteristics of these filters allow for effective filtration with only thin filter layers being required. The process of filtering air utilizing these filters is also disclosed.

Detection Of Antibodies To Squalene Iii. Naturally Occurring Antibodies To Squalene In Humans And Mice, Gary R. Matyas, Mangala Rao, Phillip R. Pittman, Robert Burge, Iris E. Robbins, Nabila M. Wassef, Brandie Thivierge, Carl R. Alving

United States Army: Publications

An ELISA-based assay is described for the measurement of antibodies to squalene (SQE) in human serum and plasma. The assay was adapted from the previously described assay for murine antibodies to SQE (J. Immunol. Methods 267 (2002) 119). Like the murine SQE antibody assay, the human antibody assay used sterile cell culture 96-well plates coated with SQE (20 nmol/well). Phosphate-buffered saline (PBS)–0.5% casein was used as both a blocking agent and dilution buffer. The assay has a high through-put capacity and is reproducible and quantitative. This assay was used to evaluate samples from three different human cohorts. The first cohort …

Heterotrophic Plate Count Bacteria—What Is Their Significance In Drinking Water?, Martin J. Allen, Stephen C. Edberg, Donald J. Reasoner

United States Environmental Protection Agency: Publications

While the literature documents the universal occurrence of heterotrophic plate count (HPC) bacteria in soils, foods, air, and all sources of water, there is a lingering question as to whether this group of organisms may signal an increased health risk when elevated populations are present in drinking water. This paper reviews the relevant literature on HPC bacteria in drinking water, the lack of clinical evidence that elevated populations or specific genera within the HPC flora pose an increased health risk to any segment of the population, and the appropriate uses of HPC data as a tool to monitor drinking water …

Laboratory Automation Curriculum At Keck Graduate Institute, James D. Sterling

Business and Information Technology Faculty Research & Creative Works

A novel curriculum in laboratory automation and high-throughput technologies has been developed at the Keck Graduate Institute (KGI) over the past five years as part of the professional master's degree program in applied life sciences. The goal of this curriculum has been four-fold: (1) motivate study by describing the need for automation through several example problems in combinatorial biological discovery, (2) provide elements of fundamental engineering science that are required for the development of the technologies and tools that enable automation, (3) provide opportunities for the students to see and use state-of-the-art instruments, learn about existing industry standards, and to …

Photo-Control Of Dna Oligonucleotides With Cage Compounds, Bilal Ghosn

LSU Master's Theses

Many biochemical processes in which DNA and other nucleic acids participate are central to functions in both living cells and in molecular biology assays. While many compounds have been used to regulate the activity of DNA, these strategies are limited to the aqueous-based diffusion of the activator to the target DNA molecule. An improvement to the induction of DNA bioactivity is to move to a light-based modulation. This research demonstrates a light-based technique using a photo-cleavable cage compound to transiently inactivate DNA hybridization. Function can be restored with exposure to near-UV light, allowing for temporal control of DNA oligonucleotide (ODN) …

Progress In The Development Of A Recombinant Vaccine For Human Hookworm Disease: The Human Hookworm Vaccine Initiative, Peter J. Hotez, Bin Zhan, Jeffrey M. Bethony, Alex Loukas, Angela Williamson, Gaddam Narsa Goud, John M. Hawdon, Azra Dobardzic, Reshad Dobardzic, Kashinath Ghosh, Maria Elena Bottazzi, Susana Mendez, Bernard Zook, Yan Wang, Sen Liu, Idong Essiet-Gibson, Sophia Chung-Debose, Shuhua Xiao, David Knox, Michael Meagher, Mehmet Inan, Rodrigo Correa-Oliviera, Paul Vilk, Herman R. Shepherd, Walter Brandt, Philip K. Russell

Michael Meagher Publications

Hookworm infection is one of the most important parasitic infections of humans, possibly outranked only by malaria as a cause of misery and suffering. An estimated 1.2 billion people are infected with hookworm in areas of rural poverty in the tropics and subtropics. Epidemiological data collected in China, Southeast Asia, and Brazil indicate that, unlike other soil-transmitted helminth infections, the highest hookworm burdens typically occur in adult populations, including the elderly. Emerging data on the host cellular immune responses of chronically infected populations suggest that hookworms induce a state of host anergy and immune hyporesponsiveness. These features account for the …

A Value Focused Thinking Approach To Software Interface In A Complex Analytical Domain, Christopher M. Mcgee

Theses and Dissertations

The intelligence community is faced with an extensive amount of data. Software programs are being developed to examine this issue of data overload and to develop solutions. The responsibility of making the final software decision lies on the analyst, therefore, the interface is the key to linking the intelligence data to the processing and results, If the interface is difficult and complex, the software will be less likely to be used. A methodology must be created which can objectively evaluate the effectiveness of the interface, This methodology will also measure the improvements in the interface's effectiveness that result when various …

Pichia Pastoris Fermentation With Mixed-Feeds Of Glycerol And Methanol: Growth Kinetics And Production Improvement, Wenhui Zhang, Karen J. Hywood Potter, Bradley A. Plantz, Vicki L. Schlegel, Leonard A. Smith, Michael M. Meagher

Papers in Biochemical Engineering

Fed-batch fermentation of a methanol utilization plus (Mut+) Pichia pastoris strain typically has a growth phase followed by a production phase (induction phase). In the growth phase glycerol is usually used as carbon for cell growth while in the production phase methanol serves as both inducer and carbon source for recombinant protein expression. Some researchers employed a mixed glycerol-methanol feeding strategy during the induction phase to improve production, but growth kinetics on glycerol and methanol and the interaction between them were not reported. The objective of this paper is to optimize the mixed feeding strategy based on growth kinetic studies …

Flight Physician - January, 2003, Civil Aviation Medical Association

Browse all Civil Aviation Medical Association Newsletters

A twenty page newsletter of the Civil Aviation Medical Association. The newsletter provided news about civil aviation medicine and information related to the organization.

Scale-Up Of The Fermentation And Purification Of The Recombinant Heavy Chain Fragment C Of Botulinum Neurotoxin Serotype F, Expressed In Pichia Pastoris, Scott K. Johnson, Wenhui Zhang, Leonard A. Smith, Karen J. Hywood-Potter, S. Todd Swanson, Vicki L. Schlegel, Michael M. Meagher

Michael Meagher Publications

A recombinant heavy chain fragment C of botulinum neurotoxin serotype F (BoNTF(Hc)) has been expressed in Pichia pastoris for use as an antigen in a proposed human vaccine. P. pastoris cells were grown using glycerol batch, glycerol fed-batch, and methanol fedbatch methods to achieve high cell densities. The total cellular protein recovered after homogenization was 72 mg/g of cell paste. BoNTF(Hc) was purified from soluble Pichia cell lysate employing ion-exchange chromatographic (IEC) and hydrophobic interaction chromatographic (HIC) methods developed at the bench scale using 10–100mL columns. The process was performed at the pilot scale using 1–4 L columns for evaluation …

Novel Small Molecule Inhibitors Of Botulinum Neurotoxin A Metalloprotease Activity, James C. Burnett, James J. Schmidt, Robert G. Stafford, Rekha G. Panchal, Tam L. Nguyen, Ann R. Hermone, Jonathan L. Vennerstrom, Connor F. Mcgrath, Douglas J. Lane, Edward A. Sausville, Daniel W. Zaharevitz, Rick Gussio, Sina Bavari

United States Army: Publications

Botulinum neurotoxins (BoNTs) are among the most lethal biological substances to have been weaponized and are listed as biodefense category A agents. Currently, no small molecule (non-peptidic) therapeutics exist to counter this threat; hence, identifying and developing compounds that inhibit BoNTs is a high priority. In the present study, a high-throughput assay was used to identify small molecules that inhibit the metalloprotease activity of BoNT serotype A light chain (BoNT/A LC). All inhibitors were further verified using a HPLC-based assay. Conformational analyses of these compounds, in conjunction with molecular docking studies, were used to predict structural features that contribute to …

Protracted Protection To Plasmodium Berghei Malaria Is Linked To Functionally And Phenotypically Heterogeneous Liver Memory Cd8+ T Cells1, Dmitri Berenzon, Robert J. Schwenk, Lisa Letellier, Mimi Guebre-Xabier, Jackie Williams, Urszula Krzych

United States Army: Publications

We previously demonstrated that protection induced by radiation-attenuated (y) Plasmodium berghei sporozoites is linked to MHC class I-restricted CD⁸⁺ T cells specific for exoerythrocytic-stage Ags, and that activated intrahepatic memory CD⁸⁺ T cells are associated with protracted protection. In this study, we further investigated intrahepatic memory CD⁸⁺ T cells to elucidate mechanisms required for their maintenance. Using phenotypic markers indicative of activation (CD44, CD45RB), migration (CD62L), and IFN-y production, we identified two subsets of intrahepatic memory CD⁸⁺ T cells: the CD44^highCD45RB^lowCD62L^lowCD122^low phenotype, representing the dominant effector …