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Tattletales And T-Bow Update 20141018, George McNamara 2014 The University of Texas

Tattletales And T-Bow Update 20141018, George Mcnamara

George McNamara

Tattletales and T-Bow Update 20141018

http://works.bepress.com/gmcnamara/63

Tattletales is my concept to multiplex fluorescent protein biosensors ("Tattletales", which is also the overall concept name) and multicolor FP reporters ("T-Bow" for Rainbow T-cells and tumor cells) to study the physiology of live cells.

These are based on:

1. LacI-GFP :: LacO operator synthetic tandem repeat array (Robinett et al 1996 JCB, Fig 4A, http://jcb.rupress.org/content/135/6/1685.full.pdf )

2. the plethora of fluorescent protein biosensors - see page 1 of this "63" download site.

3. the many colors, and combinations of colors (CY11.5 ...


Validating Tbtif2-Interacting Candidates, Fan Wu, Jennifer Berkey, Joshua Hellsing, Nisha Thaker, Bibo Li 2014 Cleveland State University

Validating Tbtif2-Interacting Candidates, Fan Wu, Jennifer Berkey, Joshua Hellsing, Nisha Thaker, Bibo Li

Undergraduate Research Posters 2014

Transmitted by the tsetse fly, Trypanosoma brucei is a protozoan parasite that causes sleeping sickness in human and nagana in cattle. While infecting the bloodstream and central nervous system, T. brucei evades the immune system by altering its major surface antigen, Variant Surface Glycoproteins (VSGs), which forms a thick coat on its cell membrane. The expression sites for VSGs are at the sub-telomeric regions of T. brucei chromosomes. Telomeres, DNA-protein complexes located at the end of chromosomes, provide chromosome stability by preventing degradation of the chromosome ends. The telomere complex also regulates the sub-telomeric VSG expression and switching in T ...


Understanding Recurrent Disease: A Dynamical Systems Approach, Wenjing Zhang 2014 Western University

Understanding Recurrent Disease: A Dynamical Systems Approach, Wenjing Zhang

University of Western Ontario - Electronic Thesis and Dissertation Repository

Recurrent disease, characterized by repeated alternations between acute relapse and long re- mission, can be a feature of both common diseases, like ear infections, and serious chronic diseases, such as HIV infection or multiple sclerosis. Due to their poorly understood etiology and the resultant challenge for medical treatment and patient management, recurrent diseases attract much attention in clinical research and biomathematics. Previous studies of recurrence by biomathematicians mainly focus on in-host models and generate recurrent patterns by in- corporating forcing functions or stochastic elements. In this study, we investigate deterministic in-host models through the qualitative analysis of dynamical systems, to ...


Transcriptional Regulation Of Peptidylarginine Deiminase Type Iv: Implications For Rheumatoid Arthritis, Ali Abbas 2014 Western University

Transcriptional Regulation Of Peptidylarginine Deiminase Type Iv: Implications For Rheumatoid Arthritis, Ali Abbas

University of Western Ontario - Electronic Thesis and Dissertation Repository

High titers of anti-citrullinated protein antibodies have been detected in sera of rheumatoid arthritis (RA) patients, implicating citrullinating enzymes in the pathogenesis of RA. Peptidylarginine deiminase type IV (PAD4) is a member of the PAD family of enzymes that catalyze the post- translational modification of arginine to citrulline and has been linked with RA. However, little is known about its transcriptional regulation. Therefore, our aim was to determine how transcription of PAD4 is activated in the myeloid lineage. Using bioinformatics, a potential nuclear factor kappa B (NF-kB) binding site was identified on the PAD4 promoter. Luciferase assays were used to ...


Detection Of Microbial Catecholamines In A Model Wound Environment, Ryan F. Wasurick 2014 California Polytechnic State University

Detection Of Microbial Catecholamines In A Model Wound Environment, Ryan F. Wasurick

STEM Teacher and Researcher (STAR) Program Posters

Chronic wounds are defined as an injury that does not heal in a predictable pattern or time frame. The healing process is composed of specific stages and chronic wounds have been interrupted in one or more stages of this process. This project focuses on the bacterial component of an infected wound, specifically on the catecholamines influencing the bacteria in the wound site. The assay developed in this experiment is designed to optimally grow bacteria in a simulated eukaryotic wound environment, and evaluate biofilm expression and catecholamine production of the bacteria. Bacteria used in this study are Staphylococcus aureus and Pseudomonas ...


Potential Roles Of Peroxidases In Caenorhabditis Elegans Innate Immunity, George R. Tiller, george r. tiller 2014 Texas Medical Center Library

Potential Roles Of Peroxidases In Caenorhabditis Elegans Innate Immunity, George R. Tiller, George R. Tiller

UT GSBS Dissertations and Theses (Open Access)

The production of ROS (reactive oxygen species) in response to pathogen detection is a rapid, nonspecific response that is evolutionarily conserved from nematodes to humans. ROS serve as direct and indirect effectors of innate and adaptive immunity. In Caenorhabditis elegans, a ROS burst is observed during infection and is mediated by the dual oxidase BLI-3, which produces H2O2. RNAi (RNA interference) to reduce the amount of BLI-3 results in a significant increase in susceptibility to pathogens, suggesting BLI-3 has a role in the immune response. However, H2O2 by itself is not a potent antimicrobial ...


The Role Of The C5a Receptor In Host Defense Against Listeria Monocytogenes, Daniel Calame 2014 Texas Medical Center Library

The Role Of The C5a Receptor In Host Defense Against Listeria Monocytogenes, Daniel Calame

UT GSBS Dissertations and Theses (Open Access)

Listeria monocytogenes (Lm) is a major cause of mortality resulting from food poisoning in the United States. While the complement component C5 is known to be protective in listeriosis, it is unknown how its cleavage fragment C5a participates. Here we show in a model of systemic Lm infection that the C5a receptor is essential for host defense. C5aR-/- mice have reduced survival and increased bacterial burden in the liver and spleen in comparison to WT mice. Surprisingly, C5aR-/- mice also have a dramatic reduction in splenocyte numbers resulting from elevated cell death as indicated by TUNEL staining and caspase 3 ...


High Fat Diet Feeding Results In Gender Specific Steatohepatitis And Inflammasome Activation, Michal Ganz, Timea Csak, Gyongyi Szabo 2014 University of Massachusetts Medical School

High Fat Diet Feeding Results In Gender Specific Steatohepatitis And Inflammasome Activation, Michal Ganz, Timea Csak, Gyongyi Szabo

Gastroenterology Publications and Presentations

AIM: To develop an animal model that encompasses the different facets of non-alcoholic steatohepatitis (NASH), which has been a challenge.

METHODS: In this study, we used a high fat diet (HFD) feeding supplemented with fructose and sucrose in the water mimicking the high-fructose corn syrup that is abundant in the diet in the United States. We used C57Bl/6 wild-type mice for short and long-term feedings of 6 and 16 wk respectively, and evaluated the extent of liver damage, steatosis, and inflammasome activation. Our methods included histopathological analysis to assess liver damage and steatosis, which involved H and E and ...


The Hot Zone And The Cold War, Peter Nichols 2014 Colby College

The Hot Zone And The Cold War, Peter Nichols

Colby Magazine

It was a time when, in the nation's schools, drills for nuclear war were as frequent as fire drills- when Americans and their Soviet counterparts lived with a constant fear of imminent worldwide destruction. "You had a cultural mindset that was apocalyptic," said Robert S. Weisbrot, Christian A. Johnson Distinguished Teaching Professor of History at Colby, "that believed you could not guarantee that your wonderful suburban home and family and community...would survive ten more seconds."

"It was not simply a matter of two countries that are adversaries building up weapons," said Weisbrot. "Each saw the other as the ...


American Society Of Parasitologists Newsletter, V. 36, Nos. 1-2, Spring-Summer 2014, Scott Lyell Gardner 2014 University of Nebraska - Lincoln

American Society Of Parasitologists Newsletter, V. 36, Nos. 1-2, Spring-Summer 2014, Scott Lyell Gardner

Newsletter of the American Society of Parasitologists

Includes editor's note and contents, a report to the ASP by President Janovy, information about a new parasitology book, a report of the ASP Meeting in New Orleans, information about the transfer of the U.S. National Parasite Collection from the Agricultural Research Service to the Smithsonian Institution, and a list of credits and ASP Affiliates.


Understanding The Factors Affecting Microbiological Quality Of Wheat Milled Products: From Wheat Fields To Milling Operations, Luis E. Sabillón Galeas 2014 University of Nebraska - Lincoln

Understanding The Factors Affecting Microbiological Quality Of Wheat Milled Products: From Wheat Fields To Milling Operations, Luis E. Sabillón Galeas

Dissertations & Theses in Food Science and Technology

Although regarded as a low-risk commodity, wheat flour-based mixes have been implicated in several food safety incidents. The present thesis is a compilation of five scientific manuscripts on the effect of weather variation, milling steps and implementation of pre-milling interventions on the microbiological quality and safety of wheat and milled products. The first manuscript is a review of the microbiological quality and safety of wheat-based products. Despite the low water activity, wheat flour may harbor dormant but viable microorganisms, which could lead to safety concerns when flour is used to produce refrigerated dough products. The second manuscript illustrates the effect ...


Fty720 (Fingolimod) Provides Insight Into The Molecular Mechanisms Of Multiple Sclerosis, Madelyn Elizabeth Crawford 2014 University of Tennessee, Knoxville

Fty720 (Fingolimod) Provides Insight Into The Molecular Mechanisms Of Multiple Sclerosis, Madelyn Elizabeth Crawford

Pursuit - The Journal of Undergraduate Research at the University of Tennessee

Multiple sclerosis (MS) is a neurodegenerative disorder caused by a prolonged immune- mediated inflammatory response that targets myelin. Nearly all of the drugs approved for the treatment of MS are general immunosuppressants or only function in symptom management. The oral medication fingolimod, however, is reported to have direct therapeutic effects on cells of the central nervous system in addition to immunomodulatory functions. Fingolimod is known to interact with sphingosine-1-phosphate (S1P) receptors, and the most widely- accepted theory for its mechanism of action is functional antagonism of the receptor. This review examines significant neuromodulatory effects achieved by functional antagonism of the ...


Post-Apartheid South Africa’S Ultimate Challenge, Nahomie Julien 2014 Georgia State University

Post-Apartheid South Africa’S Ultimate Challenge, Nahomie Julien

DISCOVERY: Georgia State Honors College Undergraduate Research Journal

Throughout time, South Africa has experienced many upheavals, be they slavery and apartheid or natural, socioeconomic, and political misfortunes.Just after overcoming the oppression of Apartheid, South Africans have to face one of the deadliest illness in the world: HIV/AIDS. HIV/AIDS is currently the number one killer in South Africa(WHO, 2012). This pandemic further worsen the struggles of the nation, obstructing its educational, financial, and political recovery (Oglethorpe, & Gelman, (2008).; Weiser, et al., 2007.). This paper seeks to analyze how apartheid—or rather, its demise—contributed to the alarmingly rapid spread of this pandemic in South Africa. In so ...


Sub-Inhibitory Antibiotics Enhance Virulence, Persistence, And Pathogenesis Of Uropathogens, Lee W. Goneau 2014 Western University

Sub-Inhibitory Antibiotics Enhance Virulence, Persistence, And Pathogenesis Of Uropathogens, Lee W. Goneau

University of Western Ontario - Electronic Thesis and Dissertation Repository

In addition to their bactericidal effects, antibiotics are potent signal mediators at sub-inhibitory levels in the environment. The ability to modulate community structure in this niche raises concerns over their capacity to influence pathogenesis in patients during antibiotic therapy. This concept forms the basis of this thesis, and is explored using models of prophylactic therapy for recurrent urinary tract infection (UTI) management.

Sub-inhibitory ciprofloxacin, ampicillin, and gentamicin were found to augment virulence in vitro, increasing adherence and urothelial cell invasion in uropathogenic Escherichia coli (UPEC) and Staphylococcus saprophyticus. In addition, biofilm formation was increased, and swarming motility decreased. In UPEC ...


Tattletales And T-Bow Update 20140602mon, George McNamara 2014 The University of Texas

Tattletales And T-Bow Update 20140602mon, George Mcnamara

George McNamara

Tattletales and T-Bow Update 20140602Mon

http://works.bepress.com/gmcnamara/42

Please see also http://works.bepress.com/gmcnamara/26

Tattletales: multiplex fluorescent protein biosensors by spatial localization with TALE-FPs, Cas9-FPs, ZF-FPs, LacI-FPs, TetR-FPs, etc.

T-Bow: Rainbow T-cells and Tumor cells (and ES cells, iPS cells, other cells and organisms). You can think of this as "Brainbow meets TALENs/Cas9/ZFNs/other DNA sequence specific binding proteins".

If not familiar with Brainbow, see

http://en.wikipedia.org/wiki/Brainbow

If not familiar with TALENs, Cas9, etc, see

http://www.addgene.org/genome_engineering/

Big idea: localizing fluorescent proteins - and/or Nano-Lanterns ...


Severity Of Infectious Mononucleosis (Im) Correlates With The Frequency Of Crossreactive Influenza A Virus (Iav)-M1 And Epstein Barr Virus (Ebv)-Bmlf-1-Specific Cd8 T Cells, Nuray Aslan, Levi B. Watkin, Anna Gil, Katherine Luzuriaga, Liisa K. Selin 2014 University of Massachusetts Medical School

Severity Of Infectious Mononucleosis (Im) Correlates With The Frequency Of Crossreactive Influenza A Virus (Iav)-M1 And Epstein Barr Virus (Ebv)-Bmlf-1-Specific Cd8 T Cells, Nuray Aslan, Levi B. Watkin, Anna Gil, Katherine Luzuriaga, Liisa K. Selin

UMass Center for Clinical and Translational Science Research Retreat

During EBV-associated IM IAV-specific crossreactive memory T cells are activated and play a role in disease severity. In HLA-A2+ IM patients, influenza M158 (IAV-M1)-specific CD8 memory T cell responses crossreacted with two different EBV lytic epitopes, BMLF1280 (17/29) and BRLF1190 (19/20). Furthermore, 11/22 IM patients demonstrated some intra-viral crossreactivity between EBV-BRLF1 and -BMLF1 responses. Disease severity of IM directly correlated with significantly increased frequencies of crossreactive IAV-M1/EBV-BMLF1, IAV-M1, and EBV-BMLF1 specific CD8 cells, and with mean viral load over the first 5 weeks of infection. Disease severity did not correlate with BRLF1 or M1/BRLF1 ...


Acute Symptomatic Influenza A Virus (Iav) Infection In Humans Leads To Expansion Of Highly Diverse Cd8 T Cell Repertoires Crossreactive With Persistent Epstein Barr Virus (Ebv), Anna Gil, Rabinarayan Misha, Nuray Aslan, Liisa K. Selin 2014 University of Massachusetts Medical School

Acute Symptomatic Influenza A Virus (Iav) Infection In Humans Leads To Expansion Of Highly Diverse Cd8 T Cell Repertoires Crossreactive With Persistent Epstein Barr Virus (Ebv), Anna Gil, Rabinarayan Misha, Nuray Aslan, Liisa K. Selin

UMass Center for Clinical and Translational Science Research Retreat

The competence of T cell responses predominantly depends on how efficient T cell receptors (TCRs) are at recognizing antigenic epitopes. We show here that during acute severely symptomatic IAV infection there was an expansion of IAV-M1/EBV-BRLF1 and IAV-M1/EBV-BMLF1 double-tetramer+ cells directly ex-vivo in 5 HLA-A2+ patients. We questioned whether this expansion specific to these two different crossreactive responses would lead to alterations in the TCR repertoire of the IAV-M158 , EBV-BRLF1109 and -BMLF1280 from before, during and following acute IAV infection. Using staining with Vb MAb we found that T cell responses generated to these epitopes became surprisingly more ...


Therapeutic Monoclonal Antibodies To Prevent Tuberculosis Infection, Andrew Crowley, Chloe Emery, Gregory W. Martens, Megan K. Proulx, Jon D. Goguen, Christopher M. Sassetti, Colby A. Souders 2014 University of Massachusetts Medical School

Therapeutic Monoclonal Antibodies To Prevent Tuberculosis Infection, Andrew Crowley, Chloe Emery, Gregory W. Martens, Megan K. Proulx, Jon D. Goguen, Christopher M. Sassetti, Colby A. Souders

UMass Center for Clinical and Translational Science Research Retreat

Mycobacteria tuberculosis (Mtb) is a major cause of human morbidity and mortality. Transmission occurs through inhalation of aerosolized Mtb and the initial infection is believed to occur primarily in the alveolar macrophage, although Mtb can infect other cells residing in the lung including dendritic cells, pneumocytes and M cells. Several molecules derived from Mtb are involved in the attachment of the organism to host receptors (opsonic and non-opsonic), which have been reasonably well elucidated. However, a complete understanding of how Mtb attaches to the host and the relative importance of each mechanism on the outcome of infection remains elusive. We ...


Topical Anti-Tnf-Α Antibody Enhances Wound Healing In Diabetic Mice, Genevieve Broderick, Heather M. Strom, Ronald A. Ignotz 2014 University of Massachusetts Medical School

Topical Anti-Tnf-Α Antibody Enhances Wound Healing In Diabetic Mice, Genevieve Broderick, Heather M. Strom, Ronald A. Ignotz

UMass Center for Clinical and Translational Science Research Retreat

Chronic wounds such as venous leg ulcers and diabetic foot ulcers represent a significant health problem for which current treatment options are limited and not highly effective. These wounds have major impacts on the quality of life for affected individuals, frequently lead to amputation, and cost many thousands of dollars per year to treat. Assessment of wound fluid from such wounds indicates greatly elevated and persistently high levels of inflammatory cytokines, in particular Tumor Necrosis Factor-alpha (TNF-α). As such, these wounds are not in a physiologic state conducive to healing. Our study in diabetic mice (db/db) suggests that topical ...


Targeted Mutagenesis Of A Therapeutic Human Monoclonal Igg1 Antibody Prevents Gelation At High Concentrations, Paul Casaz, Elisabeth N. Boucher, Rachel Wollacott, Sadettin S. Ozturk, William D. Thomas Jr., Yan Wang 2014 University of Massachusetts Medical School

Targeted Mutagenesis Of A Therapeutic Human Monoclonal Igg1 Antibody Prevents Gelation At High Concentrations, Paul Casaz, Elisabeth N. Boucher, Rachel Wollacott, Sadettin S. Ozturk, William D. Thomas Jr., Yan Wang

UMass Center for Clinical and Translational Science Research Retreat

A common challenge encountered during development of high concentration monoclonal antibody formulations is preventing self-association. Depending on the antibody and its formulation, self-association can be seen as aggregation, precipitation, opalescence or phase separation. Here we report on an unusual manifestation of self-association, formation of a semi-solid gel or “gelation”. Therapeutic monoclonal antibody C4 was isolated from human B cells based on its strong potency in neutralizing bacterial toxin in animal models. The purified antibody possessed the unusual property of forming a firm, opaque white gel when it was formulated at concentrations >40 mg/mL and the temperature was <6oC. Gel formation was reversible and was affected by salt concentration or pH, suggesting a charge interaction between IgG monomers. However, formulation optimization could not completely prevent gelation at high concentrations so a protein engineering approach was sought to resolve the problem. A comparison of the heavy and light chain amino acid sequences to consensus germline sequences revealed 16 amino acid sequence differences in the framework regions that could be involved with gelation. Restoring the C4 framework sequence to consensus germline residues by targeted mutagenesis resulted in no gel formation at 50 mg/ml at temperatures as low as 0oC. Additional genetic analysis was used to identify the key residue(s) involved in the gelation. A single substitution in the native antibody, replacing heavy chain glutamate 23 with lysine, was found sufficient to prevent gelation, while a double mutation, replacing heavy chain serine 85 and threonine 87 with arginine, increased the temperature at which gel formation initiated. These results indicate that the temperature dependence of gelation may be related to conformational changes near the charged residues or the regions interact with. Our work provided a molecular strategy that can be applied to improve the solubility of other therapeutic antibodies.


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