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Smalls-Mantey, Sample F31 Application and Summary Statement. We are truly indebted to the grantee who allowed us to post this outstanding F31 sample application to help the next generation of quotes investigators write applications. The F31 sample application below doesn’t reflect the complex layouts of the actual SF 424 PDF forms. Instead, we’ve provided a list of key fields from chapter those forms and quotes the applicant’s responses. In addition, we redacted some applicant-specific information such as contact information, publications in press, and budget specifics. The text of this application is copyrighted. You may use it only for nonprofit educational purposes provided the document remains unchanged and the PI, the grantee organization, and NIAID are credited. Questions? Contact firstname.lastname@example.org. We also offer other outstanding Sample Applications. [Our Web version of this sample application can't reproduce the layout of the actual PDF forms.
Here is the summary that eRA Commons generated at combustion of alcohols the start of the sample app, which reflects many key fields from that form.] Project/Performance Site Locations. [Our Web version of this sample application can't reproduce the betray, layout of the actual PDF forms. Closer Look The Scenes Information Popular. Performance sites are in New York, Maryland, and the United Kingdom.] Research Related Other Project Information. [Our Web version of quotes betray this sample application can't reproduce the layout of the actual PDF forms.
Here are the of alcohols, key fields from that form and the applicant's responses.] Understanding the basis of an quotes, immune response that controls infection or provides sterilizing immunity remains a major goal in the search for define gonzo effective vaccines or immunotherapies for HIV. Antibodies (Abs) induced by candidate vaccines to the surface envelope glycoprotein have not neutralized a broad array of primary virus isolates. For this reason, eliciting a cytotoxic cellular response has been the primary goal in most recent vaccine trials. However, this approach has not been successful in containing viral replication in vaccinees that have become HIV-infected. Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. In ADCC, Fc-bearing Abs bind viral epitopes coating an infected CD4+ target T cell and an Fc receptor bearing effector, most commonly natural killer cells (NKs), bind the Ab and quotes use perforin to deliver granzymes which induce apoptosis in of alcohols the target.
We want to study ADCC in infected patients to understand the magnitude and characteristics of the best responses achieved by quotes betray, natural infection. First, we will compare ADCC mediated by the sera of a cohort of patients using a granzyme B cytotoxicity assay developed in our lab. Based on combustion these findings, we will select the quotes, sera of patients with the define gonzo, most ADCC, generate monoclonal Abs (mAbs), and characterize the mAbs based on epitope specificity, affinity, potency, breadth, IgG isotype, and Fc type. We will also evaluate whether ADCC is quotes betray, disparate from classical neutralization. Define Gonzo. Finally, we will use microscopy to examine the synapse between effectors, Abs, and targets. The outcome of this research will provide insight into the characteristics of Abs that mediate ADCC that are likely important goals in the design of HIV vaccines or immunotherapies. Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is betray, a function that has been shown to mediate protection from lentiviral infection. We hypothesize that variations in look at the Actors, Information Popular Grease ADCC activity of sera are dictated by the amount, specificity, and subclass of HIV-specific antibodies. Aim 1 : Characterize the potency of sera of HIV-infected individuals in ADCC. Aim 2 : Characterize the specificity and breadth of antibodies with ADCC activity.
Aim 3 : Characterize the structure and function of the betray, target-effector synapse using both fixed and live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM) and combustion tomography. Understanding the betray, role of antibody-dependent cellular cytotoxicity (ADCC) in Closer Information on the Film, Grease HIV could provide important insights for induction of betray this activity through vaccination. This project seeks to characterize the Abs that mediate ADCC and image the functional synapse formed by cellular components involved in ADCC with the frost analysis, goal of defining new goals for the development of HIV vaccines and quotes betray therapeutics. Gallo RC, Salahuddin SZ, Popovic M. Frequent detection and isolation of cytopathic retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS. Science. 1984; 224: 500-3.
Barre-Sinoussi F, Chermann JC, Rey F, Nugeyre MT, Chamaret S, Gruest J, et al. Isolation of a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science. 1983; 220(4599): 868-71. Organization WH.
Global summary of the define gonzo, AIDS epidemic, December 2007. Quotes. 2007 [cited 2009 September 18, 2009]; Hessell AJ, Hangartner L, Hunter M, Havenith CE, Beurskens FJ, Bakker JM, et al. Fc receptor but not complement binding is important in antibody protection against HIV. Nature. Explain Based. 2007; 449(7158): 101-4. www.hivresearh.org. RV144 Phase III HIV Vaccine Trial Press Release. 2009. Chung A, Rollman E, Johansson S, Kent SJ, Stratov I. The utility of ADCC responses in HIV infection.
Current HIV research. 2008; 6(6): 515-9. Kohl S, Starr SE, oleske JM, Shore SL, Ashman RB, Nahmias AJ. Human monocytemacrophage- mediated antibody-dependent cytotoxicity to herpes simplex virus-infected cells. J Immunol. 1977; 118(3): 729-35. Forthal DN, Landucci G. In vitro reduction of virus infectivity by antibody-dependent cellmediated immunity. Betray. Journal of immunological methods. 1998; 220(1-2): 129-38. Stratov I, Chung A, Kent SJ. Behavior. Robust NK cell-mediated human immunodeficiency virus (HIV)-specific antibody-dependent responses in HIV-infected subjects.
J Virol. 2008; 82(11): 5450-9. Gomez-Roman VR, Florese RH, Patterson LJ, Peng B, Venzon D, Aldrich K, et al. A simplified method for the rapid fluorometric assessment of antibody-dependent cell-mediated cytotoxicity. Journal of immunological methods.
2006; 308(1-2): 53-67. Migueles SA, Osborne CM, Royce C, Compton AA, Joshi RP, Weeks KA, et al. Lytic granule loading of CD8+ T cells is required for HIV-infected cell elimination associated with immune control. Quotes. Immunity. 2008; 29(6): 1009-21. Vago L, Perna SK, Zanussi M, Mazzi B, Barlassina C, Stanghellini MT, et al. Loss of mismatched HLA in leukemia after stem-cell transplantation. N Engl J Med. Closer Look At The Actors, On The Popular Grease. 2009; 361(5): 478- 88. Ziegner U, Campbell D, Weinhold K, Frank I, Rutstein R, Starr SE. Deficient antibodydependent cellular cytotoxicity against human immunodeficiency virus (HIV)-expressing target cells in perinatal HIV infection.
Clin Diagn Lab Immunol. 1999; 6(5): 718-24. Brunetta E, Fogli M, Varchetta S, Bozzo L, Hudspeth KL, Marcenaro E, et al. The decreased expression of Siglec-7 represents an early marker of dysfunctional natural killer cell subsets associated with high levels of HIV-1 viremia. Blood. 2009.
Ljunggren HG, Ohlen C, Hoglund P, Yamasaki T, Klein G, Karre K. Afferent and quotes efferent cellular interactions in natural resistance directed against MHC class I deficient tumor grafts. Behavior Example. J Immunol. Quotes. 1988; 140(2): 671-8. Of A Database. De Maria A, Ferraris A, Guastella M, Pilia S, Cantoni C, Polero L, et al. Expression of HLA class I-specific inhibitory natural killer cell receptors in HIV-specific cytolytic T lymphocytes: impairment of betray specific cytolytic functions. Proceedings of the National Academy of Sciences of the United States of America. Are No Children Chapter. 1997; 94(19): 10285-8. Ahmad R, Sindhu ST, Toma E, Morisset R, Vincelette J, Menezes J, et al. Evidence for a correlation between antibody-dependent cellular cytotoxicity-mediating anti-HIV-1 antibodies and prognostic predictors of betray HIV infection. J Clin Immunol. 2001; 21(3): 227-33.
Lambotte O, Ferrari G, Moog C, Yates NL, Liao HX, Parks RJ, et al. Analysis. Heterogeneous neutralizing antibody and antibody-dependent cell cytotoxicity responses in HIV-1 elite controllers. AIDS. 2009; 23(8): 897-906. Quotes Betray. Dalgleish A, Sinclair A, Steel M, Beatson D, Ludlam C, Habeshaw J. Failure of ADCC to predict HIV-associated disease progression or outcome in Behavior a haemophiliac cohort. Clin Exp Immunol. 1990; 81(1): 5-10.
Koup RA, Sullivan JL, Levine PH, Brewster F, Mahr A, Mazzara G, et al. Antigenic specificity of antibody-dependent cell-mediated cytotoxicity directed against human immunodeficiency virus in antibody-positive sera. J Virol. Quotes. 1989; 63(2): 584-90. Doria-Rose NA, Connors M. Antibody-secreting B cells in HIV infection. Curr Opin HIV AIDS. 2009; 4(5): 426-30. Burke B, Barnett SW. Broadening our view of protective antibody responses against HIV.
Current HIV research. 2007; 5(6): 625-41. Sattentau Q. Avoiding the explain of a database, void: cell-to-cell spread of human viruses. Nat Rev Microbiol. 2008; 6(11): 815-26. Chamow SM, Zhang DZ, Tan XY, Mhatre SM, Marsters SA, Peers DH, et al. A humanized, bispecific immunoadhesin-antibody that retargets CD3+ effectors to betray, kill HIV-1- infected cells. Combustion. J Immunol. 1994; 153(9): 4268-80.
Jolly C, Kashefi K, Hollinshead M, Sattentau QJ. HIV-1 cell to cell transfer across an Env-induced, actin-dependent synapse. Betray. J Exp Med. 2004; 199(2): 283-93. Groot F, Welsch S, Sattentau QJ. Efficient HIV-1 transmission from macrophages to T cells across transient virological synapses. Blood. Explain Two Advantages Database. 2008; 111(9): 4660-3. Martin N, Welsch S, Jolly C, Briggs JA, Vaux D, Sattentau QJ. Virological synapsemediated spread of quotes betray human immunodeficiency virus type 1 between T cells is sensitive to entry inhibition. J Virol.
2010; 84(7): 3516-27. Intellectual Support : My mentors Mark Connors and Quentin Sattentau are leading experts in summary their field and have mentored numerous people that have gone on to successful careers in science. This dual mentorship will teach me to aggressively approach science from two distinctive perspectives in my graduate career, something most PhD scientist don’t experience until their first post-doctoral experience. Quotes Betray. While at NIH I will have received extensive support of not taken analysis staff from betray Richard Siegel, Director of the NIH MD/PhD partnership program and Michael Lenardo, Director the NIH/Oxford/Cambridge Scholars Program. Combustion Of Alcohols. While at Oxford I will have the guidance of Lucinda Risius, Managing Director of the betray, Graduate Pathology Program and of Criminal Behavior Essay I will also be assigned a departmental advisor, outside of my lab, with whom I can discuss my research Progress. Educational Resources : Through the betray, FAES at NIH I have access to courses to improve lab specific skills and foundation knowledge.
I have already participated a Vaccine Development course. At Oxford I will take seminars at the Centre for Excellence in Teaching and Learning for training in define gonzo teaching skills. Quotes. I will also take courses pertinent to my research offered at the Dunn School of Pathology. Both the NIH and Oxford have libraries and IT staff that can aid me in scholarly research. Physical Resources : Both labs have sufficient funding to provided me any reagent/equipement need for define gonzo the proposed research. Lab Space – I am provided with a desk and betray personal desktop computer. Bench space is shared within the lab. Safety – In order to ensure our safety from hazardous blood samples we work under laminar flow hoods in a P2 facility with P3 practices. Robert Analysis. Samples – We receive blood samples from patients enrolled within our protocol. We also have access to the NIH blood blank for normal human blood samples. As provided by Quentin Sattentau: “Oxford University is held in quotes betray high international regard for its strong teaching and research background and its unique and personal teaching style.
Within the University the combustion, Department of Pathology has an excellent reputation for graduate teaching, and has achieved 100% success in DPhil graduation within 4 years for the past 3 years. The Department has a strong graduate support structure, in betray which each student has a principal supervisor, the possibility of a cosupervisor, a departmental advisor, and a college advisor. The supervisor(s) will generally meet with the explain based database, student multiple times a term, and each of the non-supervisory mentors will meet with the betray, candidate approximately once a term and define gonzo will file reports on the candidate’s progress. The candidate will be expected to attend a series of high profile external seminars that take place once a week in term time, and a regular series of quotes internal seminars from leading and more junior University and Departmental investigators and combustion of alcohols students. Adjoa will attend and participate in the weekly lab journal club, in betray which scientific papers are taken apart critically by members of the group, and a weekly laboratory meeting in which each lab member presents their ongoing research.
Adjoa will also be encouraged to chat to other members of the department during tea lunch and coffee breaks, a longstanding Oxford institution! The Department has a fully equipped quiet library for literature research and there other reading and writing. Adjoa will have her own writing up space close to quotes, her work bench, equipped with a portable computer and if required, an external screen. Adjoa will have full access to all core facilities, including the imaging suite with multiple light microscopes, a flow cytometry and sorting suite, an electron microscopy facility with full technical and scientific support. We also have strong connections with the biochemistry and chemistry departments and the Weatherall Institute of Molecular Medicine at based the John Radcliffe Hospital for obtaining clinical samples and expertise.” BD FACSAria IIu. BD Pathways 855. VICTOR Light luminescence plate reader. AutoMACS Pro Separator.
AutoMACS Classic Separator. Containment II/III Laboratory with 4 tissue culture hoods. Tissue culture. The laboratory has a new tissue culture facility comprised of 8 double hoods and associated incubators and quotes ancillary equipment that is at the Actors, and Behind the Scenes on the Film,, shared with three other virology groups. Virology. The department has a large, fully-equipped containment-III laboratory shared between two groups, with three tissue culture hoods and associated incubators and ancillary equipment, in which HIV-1 is grown and used to infect cells.
Flow cytometry. The department has a core flow cytometry facility with three BD analyzers (2 x FACScalibur 2 x LSR2) and two sorters. Molecular biology. The Sattentau laboratory has all facilities for molecular biology carried out under containment levels –I, -II and –III. Light microscopy. The department has 5 confocal microscopes of varying sophistication and resolution, two of which are set up for quotes live cell immunofluorescence work. Theories. The department has three recently acquired high-speed CCD cameras and associated microscopes for live cell immunofluorescence analysis. The Sattentau laboratory has a Zeiss Pascal confocal microscope and a new CCD camera set up with a Zeiss Axiovert 200 microscope for fixed and live cell work in quotes the containment-III laboratory, allowing real time analysis of live, infected cells. Electron microscopy.
The department has two transmission electron microscopes and a scanning electron microscope, all of which are maintained and run as a central service facility by combustion of alcohols, a skilled dedicated operator. Our collaborator in Oxford (Kay Grunewald) is located at the University Churchill campus, which is 4 miles from the Pathology Department. He maintains and quotes uses 3 microscopes set up for cryo-tomography as follows. 300keV FEG cryo-electron microscope FEI TF30 Helium Polara with GIF2002 imaging filter (2 x 2k CCD) 300keV FEG cryo-electron microscope FEI TF30 with 4 x 4kCCD 120keV cryo-electron microscope FEI T12 with 4 x 4k CCD. Research Related Senior/Key Person Profile. [Our Web version of explain paper based database this sample application can't reproduce the layout of the actual PDF forms. See below for the biographical sketch attachments.]
My interest in studying infectious diseases stems from the 2001 anthrax scare in the Washington D.C. area. Betray. I already had a strong interest in science but through my participation in the Gene Search Science Project at Catholic University the of alcohols, following summer I came to understand how medical research impacts treatment for routine illnesses and quotes betray also plays a role in national defense. Frost Analysis. Since middle school I have been preparing for a career in science. At Eleanor Roosevelt High School I was enrolled in the rigorous Science and Technology Program during which time in participated in the Gene Search Project and researched biochemical pathways that influence sickle cell anemia with Dr. William Winters at Howard University Hospital. This experience taught me how research and medicine are intertwined and inspired me to become a physicianscientist. At UMBC I was a member of the Meyerhoff Scholars Program and an HHMI Undergraduate Research Scholar where numerous lab experiences, seminars, rigorous classes, and multiple opportunities for scientific presentations prepared me for a career as a scientist. Betray. My lab skills and understanding of define gonzo a research career increased exponentially during my four years in Michael Summers’ lab. Quotes. Not only did I learn about protein purification, gel electrophoresis, NMR, and ITC while studying MuLV RNA packaging, but more importantly of the not taken, time and discipline necessary for quotes betray excellent research and the internal drive that keeps one focused. Working with Dr. Nikola Pavletich I learned the value of immediately organizing my data so that others could analyze my results quickly.
In the lab of Tom Cech I had the responsibility of designing a new protocol to study telomerase assembly, a skill that has enabled me to design a novel assay to define gonzo, investigate AIDS specific questions in my graduate studies. The medical education I’ve received at Columbia has given me a unique perspective with which I approach scientific questions. Under the tutelage of Mark Connors and Quentin Sattentau, leaders in their fields on nonprogressive HIV infection and viral transmission, my understanding of infectious disease and quotes my capacity to pursue further research in these fields will be greatly enhanced. In addition, the courses, medical preceptorships, presentations, teaching, and career development activities I am undertaking in Theories Essay the forthcoming years make me an betray, excellent P.I. for this grant. Academic and Professional Honors. 2003 National Urban League Scholarship 2003 UMBC Meyerhoff Scholar 2003 UMBC Honors College 2003 HHMI Undergraduate Research Scholar 2004 NAD Distinguished Youth Award 2005 UMBC Minority Access to Research Careers Scholar 2005 Abby Rockefeller Mauze Award Charitable Trust Scholar for Outstanding Achievement 2006 Goldwater Scholar 2007 UMBC Top Senior Biochemistry Major 2007 Graduated Sum Cum Laude from UMBC 2007 NIH/Oxford/Cambridge Scholar 2010 International AIDS Vaccine Conference Scholarship 2011 Keystone Symposia Underrepresented Minority Scholarship. 2004 Omicron Delta Kappa Leadership Honor Society 2004 Golden Key International Honor Society, Sophomore/Alumni Director 2007 Phi Beta Kappa.
Dey A, York D, Smalls-Mantey A, Summers MF. 2005. Closer Look Actors, Popular Film,. Composition and sequence-dependent binding of RNA to the nucleocapsid protein of Moloney murine leukemia virus. Biochemistry. 44(10), 3735-3744. Miyazaki Y, Irobalieva RN, Tolbert BS, Smalls-Mantey A, Iyalla K, Loeliger K, D'Souza V, Khant H, Schmid MF, Garcia EL, Telesnitsky A, Chiu W, Summers MF. Quotes. 2010. Structure of a conserved retroviral RNA packaging element by NMR spectroscopy and cryo-electron tomography. J Mol Biol.
404(5), 751-772. Smalls-Mantey A, Winter W. 2003. Robert Frost The Road. Biosynthesis of quotes betray 2,3-diphosphoglycerate in Erythrocytes. Poster presentation at ERHS Science Fair and Prince Georges County Science Fair. March 2003.
Smalls-Mantey A, Dey, A, Phillip C, Summers MF. 2004. Characterization of High-Affinity Nucleocapsid Protein Binding Sites Within the Moloney Murine Leukemia Virus RNA Packaging Signal. Poster presented at at the Actors, and Behind the Scenes Grease 2004 Annual Biomedical Research Conference for Minority Students, November 2004. Quotes Betray. Smalls-Mantey A, Min J, Pavletich N. 2005. Structural and combustion of alcohols Biochemical Studies of the Yeast Rad4-Rad23 Complex Bound to DNA.
Poster presented at Tri-Institutional Gateways to the Laboratory Program Symposium and 2005 Leadership Alliance. August 2005. Smalls-Mantey A, Miyazaki Y, Summers MF. 2007. Dimerization of betray Moloney Murine Leukemia Virus Nucleocapsid Protein Binding Sites Facilitates Binding.
Poster presented at 2007 UMBC Undergraduate Research and Creative Achievement Day. April 2007. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. Of Criminal Behavior Example. 2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells. Poster presented at 2010 Keystone HIV Vaccines (X5) Meeting. March 2010. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu.
2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells. Poster presented at 2010 Oxford Pathology Department Graduate Student Symposium. June 2010. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. 2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells.
Poster presented at 2010 NIH/Oxford/Cambridge Scholars Research Colloquium. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. 2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells. Poster presented at 2010 International AIDS Vaccine Conference. September 2010. Smalls-Mantey A. 2010.
Antibody-Dependent Cellular Cytotoxicity and the Control of HIV. Quotes Betray. Talk given at Frontiers in Biomedicine NIH/Oxford/Cambridge Scholars Program student presentation series. November 2010. [Outside research support redacted from sample.] A. Personal Statement Regarding Project.
Currently HIV-specific ADCC is Closer and Behind the Scenes Information Film, Grease, considered one of the very top priorities in the field of HIV-specific immunity. Our laboratory has assembled the best techniques and quotes betray reagents in the field to carry out two advantages database, this project. We have extensive experience in HIV-specific antibodies and cytotoxicity. In addition we have a long track record of training students to prepare them for a carreer in biomedical research. 1985 - 1986 Intern in Pediatrics, The Boston Floating Hospital, New England Medical Center, Boston, Massachusetts 1986 - 1988 Resident in quotes Pediatrics, The Boston Floating Hospital, New England Medical Center, Boston, Massachusetts 1988 – 1989 Chief Resident in Pediatrics, The Boston Floating Hospital, New England Medical Center, Boston, Massachusetts July 1989 - October 1989 Medical Staff Fellow, Respiratory Viruses Section, Laboratory of of a paper based database Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD October 1989 - June 1993 U.S. Public Health Service, Research Associate (C.O.), Respiratory Viruses Section, Laboratory of Infectious Diseases, National Institute of quotes Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD July 1993 - June 1994 U.S. Public Health Service, Clinical Associate, Clinical Center, National Institute of combustion Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD January 1994 - June 1994 Clinical Fellow in Infectious Diseases, The Childrens Hospital of betray Philadelphia, Philadelphia, PA. June 1994 - December 2004 U.S. Public Health Service, Medical Officer, National Institute of Allergy and Infectious Diseases, Clinical and Molecular Retrovirology Section, Laboratory of Immunoregulation, National Institutes of Health, Bethesda, MD December 2007- Present Chief, HIV-Specific Immunity Section, Laboratory of explain two advantages of a database Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of quotes Health, Bethesda, MD. Recent Honors and Editorial and Review Board Activities. 2009 - U.S.
Public Health Service Outstanding Service Medal 2009 - present Vaccine Production Development Team Scientific Advisory Board, University of Pennsylvania 2009 - present HIV Enterprise Host Genetics and HIV Diversity Working Group 2010 - present Editorial Board, Journal of Virology. C. Peer-Reviewed Publications. Migueles SA, Sabbaghian MS, Shupert WL, Bettinotti MP, Marincola FM, Schwartz D, Sullivan J, and Connors M. HLA B*5701 is highly associated with restriction of virus replication in define gonzo a subgroup of HIV infected long term nonprogressors. Quotes. PNAS 2000; 97(6): 2709-2714. Migueles SA, Laborico AC, Shupert WL, Sabbaghian MS, Rabin R, Hallahan C, Van Baarle D, Kostense S, Miedema F, McLaughlin M, Ehler L, Metcalf J, Liu S, and Connors M. HIV-specific CD8+ T cell proliferation is coupled to perforin expression and combustion of alcohols is maintained in nonprogressors. Nature Immunol 2002; 3(11); 1061-1068.
Migueles SA, Laborico AC, Imamichi H, Shupert WL, Royce C, McLaughlin M, Ehler L, Metcalf J, Liu S, Hallahan CW, and Connors M. The differential ability of betray HLA B*5701+ long-term nonprogressors and progressors to restrict HIV replication is not caused by loss of recognition of autologous viral gag sequences. J Virol 2003; 77(12); 6889-6898. Iyasere C, Tilton JC, Johnson AJ, Younes S, Yassine-Diab B, Sekaly RP, Kwok WW, Migueles SA, Laborico AC, Shupert WL, Hallahan CW, Davey RT Jr., Dybul M, Vogel S, Metcalf J, and Connors M. Diminished proliferation of HIV-specific CD4+ T Cells is associated with diminished IL-2 production and recovered by exogenous IL-2. J Virol 2003; 77(20); 10900-10909. Tilton JC, Luskin MR, Johnson AJ, Manion M, Hallahan CW, Metcalf JA, McLaughlin M, Davey RT Jr., and Connors M. Changes in frost analysis paracrine IL-2 requirement, CCR7 expression, frequency, and cytokine secretion, of HIV-specific CD4+ T cells are a consequence of antigen load. Quotes Betray. J Virol 2007; 81;2713-2725. Migueles SA, Osborne CM, Royce C, Compton AA, Joshi RP, Weeks KA, Rood JE, Berkley AM, Sacha JB, Cogliano-Shutta NA, Lloyd M, Roby G, Kwan R, McLaughlin M, Stallings S, Rehm C, O’Shea MA, Mican J, Packard B, Komoriya A, Palmer S, Wiegand AP, Maldarelli F, Coffin JM, Mellors JW, Hallahan CW, Follman DA and Connors M. Lytic Granule Loading of CD8+ T-Cells is Required for look at the and Behind Information Popular Grease HIV-Infected Cell Elimination Associated with Immune Control. Immunity 2008; 29 (6): 1009-1021.
Migueles SA, Weeks KA, Nou E, Berkley AM, Rood JE, Osborne CM, Hallahan CW, Cogliano-Shutta NA, Metcalf JA, McLaughlin M, Kwan R, Mican JM, Davey RT Jr, Connors M. Betray. Defective human immunodeficiency virus-specific CD8+ T-cell polyfunctionality, proliferation, and cytotoxicity are not restored by antiretroviral therapy. J Virol 2009 83(22): 11876-11889. Doria-Rose NA, Klein R, Daniels M, O'Dell S, Nason M, Lapedes A, Bhattacharya T, Migueles S, Wyatt RT, Korber BT, Mascola JR, Connors M. Breadth of there are no chapter HIV-Specific neutralizing activity in sera: clustering analysis and association with clinical variables. J Virol 2009 84(3): 1631-1636. Doria-Rose NA and Connors M. Quotes Betray. Antibody-secreting B cells in HIV infection. Curr Opin HIV AIDS 2009 4:426-430 Wu X, Yang ZY, Li Y, Hogerkorp CM, Schief WR, Seaman MS, Zhou T, Schmidt SD, Wu L, Xu L, Longo NS, McKee K, O'Dell S, Louder MK, Wycuff DL, Feng Y, Nason M, Doria-Rose N, Connors M, Kwong PD, Roederer M, Wyatt RT, Nabel GJ and Mascola JR. Rational design of envelope identifies broadly neutralizing human monoclonal antibodies to HIV-1.
Science 2010 329:856-861. Migueles SA and Theories of Criminal Behavior Essay example Connors M. Quotes. Long-term nonprogressive disease among untreated HIV-infected individuals: clinical implications of the road not taken analysis understanding immune control of HIV. JAMA 2010 304:194-201. Betray. [Redacted publication in press] The research direction of the HIV-Specific Immunity Section of the Laboratory of Immunoregulation is focused upon mechanisms of immunity to HIV. The laboratory is staffed by PhD and MD scientists with extensive experience in the humoral or cellular immune response to HIV. In addition we have close collaborations with investigators at the NIAID Vaccine Research Center in the area of structure-function relationships of explain two advantages of a paper based database HIV-specific antibodies. Our laboratory is quotes, well equipped with multi-laser flow cytometers, automated magnetic sorters, and imaging equipment to carry out explain two advantages of a database, this project.
Our laboratory is supported by quotes betray, the research budget of the robert frost not taken, Division of quotes betray Intramural Research. Over the past 4 years we have received supplemental funding from the frost not taken analysis, Intramural AIDS Targeted Antiretroviral Program (IATAP). Quotes Betray. This will continue for the next 2 years. Of Alcohols. We have also received funding from the Office of AIDS Research for quotes betray work on the humoral response to HIV. Positions and Employment.
1985 - 1992 Lecturer, Department of Genito-Urinary Medicine, University College London, UK 1989 - 1991 Visiting scientist, Howard Hughes Medical Institute, Columbia University, New York, NY 1992 - 1998 Staff scientist (Director of Behavior Research), Centre d’Immunologie, Marseille, France 1999 - 2003 Reader, Section of betray Infectious Disease, Imperial College London, London UK 2003 - Professor of immunology, Department of Pathology, University of Oxford, Oxford UK. Other Experience and Professional Memberships. 1995 - Member, American Association for Theories of Criminal Behavior example the Advancement of quotes Science 1998 - Member, American Society for Microbiology 1998 - Member, British Society for Immunology, UK 1998 - Member, Society for General Microbiology, UK 2005 - Member of the children, Pasteur Institute Paris France Scientific Evaluation Committee. 2005 - Member of the Vaccine Research Center (VRC) NIH Board of Scientific Councilors 2007 - Member of BMGF Vaccine Monitoring Centre SAB 2008 - Member of NIH HIVRAD/Novartis Neutralizing Antibody Vaccine Consortium SAB 2009 - Member of study section for Swedish Program for International Development 2010 - Member of NIH study section for the B cell Immunology Partnership Program for quotes betray HIV-1 Vaccine Discovery (U19) C. Selected Peer-Reviewed Publications. Most relevant to the current application. Martin N, Welsch S, Jolly C, Briggs JA, Vaux D and Sattentau QJ. (2010) Virological synapse-mediated spread of of Criminal Behavior Essay example human immunodeficiency virus type-1 between T cells is sensitive to entry inhibition. J. Virol. 84: 3516-3527. Gonzalez N, Bermejo M, Calonge E, Jolly C, Arenzana-Seisdedos F, Pablos JL, Sattentau QJ and Alcami J. (2010) SDF-1/CXCL12 production by mature dendritic cells inhibits the propagation of X4-tropic HIV-1 isolates at the dendritic cell-T-cell infectious synapse.
J Virol. 84:4341-51 Groot F, Welsch S and betray Sattentau QJ. (2008). Efficient HIV-1 transmission from macrophages to at the and Behind the Scenes Information on the Popular, T cells across transient virological synapses. Blood 111: 4660-4663. Sowinski S, Jolly CJ, Berninghausen O, Purbhoo, MA, Chauveau A, Kohler K, Oddos S, Eissmann P, Brodsky FM, Hopkins C, Onfelt B, Sattentau QJ and Davis DM (2008).
Membrane nanotubes physically connect T cells over long distances presenting a novel route for HIV-1 transmission. Quotes. Nature Cell. Biol. 10: 211-219. Jolly C and Sattentau QJ (2007).
HIV-1 assembly, budding and of alcohols cell-cell spread in T cells takes place in tetraspanin-enriched plasma membrane. J. Virol. 81:7873-7884 Jolly C and Sattentau QJ (2007). Adhesion molecule interactions facilitate human immunodeficiency virus type-1-induced virological synapse formation between T cells. J. Virol. 81: 13916-13921. Jolly C, Mitar I and betray Sattentau QJ (2007). Requirement for an intact actin and Essay tubulin cytoskeleton for efficient HIV-1 assembly and spread.
J. Virol. 81: 5547-6 Jolly C and Sattentau QJ. Human Immunodeficiency virus type-1 virological synapse formation in T cells requires lipid raft integrity. J. Quotes Betray. Virol. Are No Chapter Summary. 2005 79: 12088-12094.
Jolly C, Kashefi K, Hollinshead M and Sattentau QJ (2004) HIV-1 cell-to-cell transfer across an betray, Env-induced, actin-dependent synapse. Theories Of Criminal Behavior Essay Example. J. Exp Med. 199: 283-193. Additional recent publications of quotes important to of alcohols, the Field (in chronological order) Wegmann F, Krashias G, Luhn K, Laamanen K, Vieira S, Jeffs SA, Shattock RJ and Sattentau QJ (2011) A novel strategy for inducing enhanced mucosal HIV-1 antibody responses in quotes an anti-inflammatory environment. PlosOne 6: e15861. Behavior Essay Example. Kong L, Sheppard N, Stewart-Jones G, Robson CL, Chen H, Xu X, Krashias G, Bonomelli C, Scanlan CN, Kwong PD, Jeffs SA, Jones IM, Sattentau QJ (2010) Expression system-dependent modulation of HIV-1 envelope glycoprotein antigenicity and betray immunogenicity. J. Mol. Biol. Aug 25th epub. Krashias G, Simon K, Wegmann F, Kok WL, Ho LP, Stevens D, Skehel J, Heeney JL, Moghaddam AE and Information on the Popular Grease Sattentau QJ. (2010).
Potent adaptive immune responses induced against HIV-1 gp140 and influenza virus HA by betray, a polyanionic carbomer. Vaccine 28: 2482-2489. Zanetti G, Briggs JAG, Grunewald K, Sattentau QJ and at the and Behind Information on the Film, Fuller SD. SIV spike glycoprotein structure in situ determined by cryo-electron tomography. Plos Pathogens August 2 (8): e83 Sheppard NC, Bates AC and Sattentau QJ (2007). A functional human IgM response to quotes, HIV-1 Env after immunization with NYVAC-HIV C. AIDS 19: 524-527. [Outside research support redacted from sample.] PHS Fellowship Supplemental Form, Research Plan. [Our Web version of this sample application can't reproduce the layout of the actual PDF forms. The attachments are included below. In the original version, these attachments met the page limits.]
Understanding the basis of an immune response that controls infection or provides sterilizing immunity remains a major goal in the search for effective vaccines or immunotherapies for HIV. Theories Essay. Antibodies (Abs) induced by candidate vaccines to the surface envelope glycoprotein have not neutralized a broad array of primary virus isolates. For this reason, eliciting a cytotoxic cellular response has been the primary goal in betray most recent vaccine trials. However, this approach has not been successful in containing viral replication in vaccinees that have become HIV-infected. Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. Combustion. In ADCC, Fc-bearing Abs bind viral epitopes coating an infected CD4+ target T cell and an Fc receptor bearing effector, most commonly natural killer cells (NKs), bind the quotes betray, Ab and use perforin to deliver granzymes which induce apoptosis in the target. We want to study ADCC in of alcohols infected patients to understand the magnitude and quotes characteristics of the best responses achieved by natural infection. First, we will compare ADCC mediated by the sera of a cohort of patients using a granzyme B cytotoxicity assay developed in our lab. Of Alcohols. Based on these findings, we will select the quotes betray, sera of patients with the frost not taken analysis, most ADCC, generate monoclonal Abs (mAbs), and characterize the mAbs based on epitope specificity, affinity, potency, breadth, IgG isotype, and Fc type.
We will also evaluate whether ADCC is disparate from classical neutralization. Finally we will use microscopy to examine the synapse between effectors, Abs, and targets. The outcome of this research will provide insight into the characteristics of betray Abs that mediate ADCC that are likely important goals in the design of HIV vaccines or immunotherapies. Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is a function that has been shown to mediate protection from lentiviral infection. We hypothesize that variations in ADCC activity of sera are dictated by combustion, the amount, specificity, and subclass of HIV-specific antibodies. Aim 1 : Characterize the potency of sera of HIV-infected individuals in ADCC. In ADCC, Abs bind viral epitopes that are presented by infected CD4+ T cells. NKs expressing an Fc receptor bind the Fc domain of the Ab and use perforin to deliver granzymes to the HIV-infected cell.
Subsequently, granzymes induce apoptosis within the cell. Quotes Betray. Our lab has developed a flow cytometric assay that measures granzyme B delivered to an HIV-infected CD4+ target T cell. We will classify ADCC by combustion, the percent of target cells receiving granzyme and the elimination of targets as defined by residual percent of targets expressing p24, HIV capsid. Compare the betray, serum of HIV+ individuals with various rates of progression and viral loads to two advantages of a database, determine which contain Abs capable of mediating the highest levels of ADCC. Compare the ADCC and neutralizing activity of patient sera. Aim 2 : Characterize the specificity and breadth of antibodies with ADCC activity. Our laboratory has panels of NAbs derived from patients with known serum neutralizing or ADCC-mediating activity. Determine whether recognition of quotes betray specific epitopes is are no children chapter, required for ADCC. Define the breadth of the polyclonal sera by its ability to quotes, mediate ADCC in define gonzo CD4+ T cells infected by different clades of HIV.
Titer serum total IgG, IgG1, and IgG3 binding infected CD4+ T cells. Aim 3 : Characterize the structure and function of the target-effector synapse. Using both fixed and live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM) and betray tomography, we will examine the synapse formed between NK and other cells with potential ADCC activity (macrophages and neutrophils) and infected target cells. We will specifically investigate: The structure of a functional ADCC synapse. The kinetics of combustion ADCC function in real time and its relation to quotes betray, antibody type and specificity. A role for Theories of Criminal Behavior example antibody-dependent cell-mediated phagocytosis (ADCP) in elimination of HIV-infected cells. Quotes Betray. d. Receptors and look and Behind Information on the Film, effector molecules central to ADCC activity against HIV infected cells. Since being identified as the causative agent of quotes betray AIDS in 1983, HIV infection has reached epidemic proportions with an estimated 2 million people worldwide dying from two advantages of a database AIDS each year 1- 3. There is more public awareness about how to prevent HIV infection but with the growing incidence of HIV infection and the limitations of current antiretroviral therapy, there is an imperative need for a vaccine.
One of the most critical areas of research on vaccines for HIV is the search for correlates of immunity. At present the only method for determining the potential benefits of a vaccine with regard to protection from quotes betray infection or lowering of viral load upon infection remains very large clinical efficacy trials. Determining correlates of immunity would tremendously accelerate the development process by permitting some estimation of clinical efficacy prior to a Phase III trial. Reliable correlates would permit screening of vaccine candidates at a much earlier point in development with smaller numbers of participants. Over the past 20 years, research into correlates of immunity to HIV have largely focused on CD8+ cytotoxic T lymphocytes (CTLs) or neutralizing antibodies (NAbs). In addition to neutralization, antibodies (Abs) may fight viral infections by antibody-dependent cell-mediated phagocytosis (ADCP) or by antibody-dependent cellular cytotoxicity (ADCC). In ADCC, Abs bind viral epitopes presented on infected target cells and robert not taken analysis Fc? receptor-bearing effector cells such as natural killer cells (NKs) recognize and bind the Fc? region of these Abs. Upon cross-linking of the Fc? receptor, NKs release perforin that punctures the target’s cell membrane and deliver proteases such as granzyme B (GrB) which induce apoptosis. ADCC has become a major focus within the field following two important studies highlighting its potential importance in providing immunity to HIV.
A recent study by Burton and colleagues demonstrated that neutralizing Abs with mutated Fc? receptor binding regions were less effective in betray protecting macaques from explain of a HIV whereas Abs with impaired complement binding conferred protection to macaques, suggesting that ADCC may be more important in preventing infection than neutralization or complement activation alone 4. Prior to this study, it was widely accepted that infection of a single cell would lead to integration and quotes betray chronic infection. However, because ADCC mediated effects are on virus infected cells and not free virions, this study provided indirect evidence and proof of the concept that ADCC can eliminate lentivirus-infected cells in vivo. Are No Here Chapter Summary. In addition, some recent data in betray human efficacy trials has also greatly increased interest in ADCC as an important component of vaccine induced immunity to HIV. Closer Actors, And Behind The Scenes On The Popular Film, Grease. In September 2009 data from the phase III RV144 (Thai) vaccine trial were released and showed that the prime-boost combination of ALVACR HIV and AIDSVAXR B/E reduced HIV infection. This vaccine regimen was known to not elicit a strong cellular or NAb response. It does, however, elicit binding antibodies that have suggested to the field that the protective efficacy is likely mediated through ADCC 5. Our laboratory is part of a large international effort to characterize ADCC antibodies from this trial in quotes betray order to determine whether there were higher ADCC-mediating antibodies in uninfected vaccinees compared to explain of a database, those that were infected. Although HIV-specific ADCC is now considered an extremely important part of vaccineinduced immunity several basic questions regarding the nature of an effective ADCC response have not been addressed. For example, the quotes betray, number, affinity, and Closer at the Actors, Information Popular Film, Grease specificities of antibodies necessary to mediate ADCC remains poorly understood. Our laboratory has now developed assays that can accurately measure ADCC against HIV-infected cells. Quotes. We also have assembled panels of monoclonal antibodies from individual patients that permit a characterization of of alcohols individual antibodies that mediate HIV-specific ADCC.
One of the initial goals of this project was to adapt our current killing assay to betray, provide a more robust assay for HIV-specific ADCC. Frost The Road Not Taken. Past studies of ADCC have been hampered by the availability of assays that are only quotes partially quantitative. There are four primary methods that people use to measure ADCC 6. The most prevalent is the chromium release assays (CRA), which is highly variable, does not specifically account for the mechanism of killing, and Theories Essay is insufficiently quantitative 7. Betray. The ADCVI assay incubates targets, serum, and effectors for 24 hours and the supernatant is applied to the B95-8 cell line for 5-7 days. The endpoint is reduction of viral infection of the Theories of Criminal Behavior Essay, B95-8 cells 8. This method measures inhibition of viral production and subsequent infection but its disadvantages lie in betray the length of the explain two advantages based, assay and betray failure to measure killing of infected cells. Another assay is the intracellular cytokine staining ADCC (ICS-ADCC) assay measures IFN-?, the primary cytokine responsible for the antiviral activity of many cells including NKs, CTLs, Th1, and dendritic cells 9. The advantages of define gonzo this assay are that multiple effector functions can be measured and quotes all cells and Abs are from the same patient, but it is still a proxy that does not measure HIV-infected cell killing.
The rapid and fluorometric antibody-dependent cellular cytotoxicity (RFADCC) assay directly measures cell killing but requires whole protein 10. While use of protein pulsed targets is define gonzo, a sensitive method, it likely places artificially high amounts of protein on the surface of infected cells, potentially overestimating the efficacy of ADCC. Recently our lab has developed a highly quantitative flow cytometric assay to measure GrB cytotoxicity using primary HIV-infected CD4+ T cells as targets and autologous NKs as effectors. This is an betray, adaptation of an explain two advantages of a based, assay for measuring the cytotoxic activity of CTLs developed in our lab11. This assay measures the cleavage of a GrB substrate, and eliminates background cell death using a Live/Dead stain to quotes betray, produce accurate quantitative and highly reproducible results. Look Actors, Information Popular Film,. In brief, CD4+ T cells are stimulated for 3 days and HIV infected. Autologous, negatively selected NKs are used as effectors. Quotes. Effectors and targets are labeled with surface dyes so that they can be discriminated. NKs, targets and serum are incubated in the presence of a granzyme cleavable fluorescent substrate.
Granzyme delivery to the target cell and Essay elimination of HIV-infected targets can then be measured by flow cytometry (Figure 1). Cellular GrB content. GrB cleaves GranToxiLux substrate (FITC), indicating the amount of GrB present within a target cell (Pacific Blue). Elimination of HIVSF162-infected cells. Live targets are stained for intracellular HIV capsid, p24 (PE). Some infected cells show downregulation of CD4 (PE-Cy7). Betray. A reduction in the number of p24 stained cells is represented as Infected Cell Elimination (ICE) and is an indication of cell death. In the look Actors, Popular Film,, example above, ICEInfected+Effectors+Serum = [(58.2-25.8)/58.2] x 100 = 54.9%.
There are several advantages to this assay over quotes betray older assays that use mismatched cell lines. Closer At The Actors, And Behind Popular. Because NKs lyse target cells in the context of MHC mismatch and quotes cause nonspecific killing, autologous cells are used in our assay to reduce background. Of A Based. 12. In addition, because NK function is quotes betray, diminished in HIV-infected patients, we use the NK cells and CD4+ T cell targets from uninfected patients in our assay as a readout for explain two advantages based the effectiveness of a given serum sample in mediating ADCC 13, 14. We observed variations in NKs from normal individuals in quotes betray their ability to mediate ADCC and choose donors that provide optimal signal to children chapter, noise ratio that we will use to quotes betray, compare the two advantages, serum of HIV+ individuals (Figure 2). The targets used in betray this assay are likely closer to the in robert frost the road vivo situation given autologous HIV-infected targets are used in the place of antigen pulsed cell lines.
Other advantages include our assay’s ability to measure the mechanism of ADCC (granzyme delivery) and the outcome (death of infected cells) (Figure 2). MHC-1 is down-regulated in virally infected cells, and the loss of quotes betray MHC-mediated killer inhibitory signals permit NKs to kill a cell 15, 16. We control for this by measuring the granzyme content of targets that are incubated only with effectors and sera from HIV-uninfected volunteers. For a positive control we are using a novel anti-CD3 Ab that will mediate ADCC to all CD3-bearing targets (Figure 3). With this Ab we have already demonstrated (1) the sensitivity and dynamic range with which we are able to measure ADCC, (2) Abs mediate ADCC in a dose dependent manner, and (3) GrB delivery is proportional to cell death. Aim 1: Characterize the magnitude of ADCC in HIV infected individuals . Rational : There is considerable controversy regarding the magnitude of ADCC activity in sera from HIV+ individuals and how it correlates with disease progression.
It has been suggested that sera from elite controllers exhibit greater ADCC activity which correlates with CD4+ T cell count 17, 18 while others have sighted no correlation between ADCC and disease progression 19, 20.We will compare the serum of Theories example HIV+ individuals with various rates of progression and viral loads to quotes betray, determine which contain Abs capable of mediating the combustion of alcohols, highest levels of ADCC. We will also compare the ADCC and neutralizing activity of patient serum to evaluate whether or not ADCC and neutralization always occur together in polyclonal sera which is betray, yet to be determined 21, 22. Methods : Over the past several years, in collaboration with the Vaccine Research Center (VRC) our laboratory has focused upon the road recruiting HIV-infected patients with broadly crossneutralizing antibodies. We have extensive clinical data and neutralization data for all of the patients. With the data we generate from our ADCC assay we are able to compare multiple indicators of quotes immunity with ADCC. Expected Outcomes/Interpretation : Based on in vitro and passive transfer macaque studies, we expect LTNPs to exhibit greater ADCC than progressors. Our current knowledge attributes LTNP delayed progression to superior CTL cytotoxicity but our data may show ADCC also to be a significant factor 11. Potential Complications : We do not foresee any difficulties in accomplishing this objective. Aim 2: Characterize MAb from serum identified as having potent ADCC activity with regard to specificity. Rational/Methods : HIV-infected cells express Envelope (Env) protein on their surface which is recognized by NAbs and presumably by ADCC-mediating Abs.
We will use standard Nabs (including b12, 2F5, 2G12, 447-52D, 17B, and define gonzo VRC01) and patient derived MAbs to probe specificities required for ADCC. An ideal HIV vaccine would be able to protect against quotes, diverse virus strains so it is important to determine whether Abs raised against one clade of virus would be able to mediate ADCC in CD4+ T cells infected by different clades. We will use multiple strains of virus from clade A/E, B, and C to infect CD+ T cells and define gonzo compare killing of target cells in the presence of serum pooled from clade B HIV+ patients. IgG1 and IgG3 are the predominant IgG isotypes with roles in ADCC. It is possible that variations in ADCC activity of sera are mediated by variations in IgG subclass. We will use infected CD4+ T cells, apply sera with various levels ADCC at the same dilutions used in quotes betray our killing assay, and measure total bound IgG, IgG1, and IgG3 by flow cytometry. We believe flow cytometry is a better way of measuring titers of binding Abs as compared to ELISA because Abs will recognize all possible epitopes on the surface of an infected cell in their natural conformation. Expected Outcomes/Interpretation : Determining the association between ADCC activity and IgG subclass may provide insights into the necessity of eliciting a specific class of IgG via particular antigens and adjuvants. Potential Complications : We now have robust assays for subclass binding and of alcohols do not expect complications with this portion of the project.
Aim 3: Evaluation of the target-effector synapse. Rational : Using microscopy to study ADCC is important because it will help us define structural and kinetic parameters and will relate structure to function. Multiple mechanisms for cell-to-cell spread of HIV have been described but few have described inhibition of viral transmission or targeted cell death 23. To date there has been no study using microscopy to investigate ADCC in HIV using NKs or macrophages as effectors and CD4+ T cells as targets. In 1994 one group designed a bispecific antibody with anti-CD4 and quotes betray anti CD3 heavy chains that would redirect CD8+ T cells to kill HIV infected targets 24.
With fluorescent microscopy they observed after 1 hour of incubation that 24% targets had formed conjugates with CD8+ T cells although many more cells had lysed. At 2 hours 50 - 80% of explain two advantages paper based database targets were conjugated and undergoing lysis and by 24 hours all targets had been lysed. However, the quotes betray, bispecific antibody may not be representative of ADCC with naturally occurring HIV-specific antibodies. In addition, the structure of the NK to HIV-infected target synapse was not examined. Methods : The lab of Quentin Sattentau has focused on examining the synapse of cells involved in HIV infection. They were the first to demonstrate HIV-1 spread between T cells via virological synapses and define gonzo nanotube structures for viral transmission between CD4+ T cells 25. They have recently demonstrated that macrophages can infect CD4+ T cells every 6 hours and that cell-tocell transfer of virus is 10-fold more efficient than free virus spread 26.
To study the kinetics of quotes ADCC in real time we will use time-lapse laser scanning confocal microscopy (time-lapse LSCM). Cell populations will be negatively selected from are no children here chapter PBMCs using magnetic cell sorting. HIV-infected CD4+ T cells will be mixed with patient serum or Abs of multiple specificities (see Aim 2) and NKs to evaluate ADCC; or neutrophils/monocytes to betray, evaluate antibody-dependent cell-mediated phagocytosis (ADCP) and applied to two advantages of a, poly-L-lysine coated cover slips. Cover slips will be fixed in paraformaldehyde, quenched with NH4Cl, permeabilized with Triton-X, and quotes betray stained for granzyme B, IgG, CD4/CD16/CD14 with fluorescent probes. After mounting, images will be acquired every minute for 6 hours using a Zeiss Pascal Axivert 200M scope.
We will use transmission electron microscopy and at the and Behind the Scenes Information Film, tomography (TEM) to betray, generate higher resolution static images of the ADCC functional synapse. For TEM targets, serum, and of Criminal Behavior example effectors will be incubated for quotes approximately 1 hour, fixed in define gonzo paraformaldehyde and quotes betray glutaraldehyde, and stained with osmium tetroxide. Actors, The Scenes On The Popular Film,. Samples will be embedded in epoxy resin, cut, and imaged using a 300kV FEI Technai TF30 27. If necessary cryo-EM will also be used for structural studies with the aid of Oxford collaborator Kay Grunewald. These experiments will be completed using the pathology department’s core bioimaging facility and I will take department’s Advanced Light Microscopy and betray Electron Microscopy course in preparation for this research. Expected Outcomes : We expect to elucidate the kinetic and functional relationship between ADCC components and answer questions such as what density of Env epitopes are needed to recruit effectors, how many effectors are recruited per target. We do not foresee any difficulties in accomplishing this objective. The results of these studies will include novel MAb with possible therapeutic value; a new tool for the study of ADCC with potential utility for HIV vaccine trials; and look Information increased understanding of basic HIV immunology. In the wake of the RV144 trial, ADCC has become a major focus in the field of HIV-specific immunity. We are very well positioned to carry out these important studies because of quotes our large collection of Behavior cells, sera, and betray detailed clinical data from patients that are part of the NIAID HIV clinic, and Theories Essay example both labs’ extensive experience in this area.
Samples are collected under the quotes betray, NIAID IRB-approved protocol 02-I-0086. There Children Summary. Protections for human subjects are in quotes accordance with NIAID Division of Intramural Research policies. Phlebotomy may cause some discomfort, and occasionally some bleeding or bruising at the site. On rare occasions, people faint while having blood drawn. Blood drawing will not exceed 150 milliliters (30 teaspoonfuls) on any day or visit. In the course of two advantages based database any time frame during this study, the total amount of blood will not exceed 10.5mL/kg, or 550 milliliters over any 8-week period, which is within the safety guidelines for blood drawing practiced at the Clinical Center. Like standard phlebotomy, leukapheresis is occasionally associated with local bleeding and bruising at the site of needle insertion.
Lightheadedness can also occur, and people faint on rare occasions. Occasional tingling in the arms and legs can occur with the procedure and can usually be relieved by chewing a calcium-rich tablet that the quotes betray, staff will provide. In addition, on very rare occasions (less than 1 in every 1,000 times), malfunction of the of Criminal Behavior Essay, leukapheresis machine can result in blood loss of up to 450 milliliters. Trained medical personnel who are equipped to deal with any of these potential problems will always be on hand during the procedure. Inclusion of Women and Minorities. Gender, Ethnicity and quotes betray Race Consideration. This study involves techniques and procedures that are currently approved for various indications. Male and female adult patients (18 years of age or older) will be enrolled without preference to gender, race or ethnicity from the pool of patients that receive care in Clinics 8 at define gonzo the NIH Clinical Center. Patients will be recruited from outside the quotes, NIH community by advertisements posted in the Clinical Center, the NIAID web site, and Behavior Essay example advertised in other media. We will attempt to facilitate minority recruitment using existing NIAID community outreach efforts involving local clinics in quotes the Washington, DC metropolitan area. In adult HIV-infected patients planning to interrupt HAART, patients will be fully counseled prior to entry into the study as to the potential risks of discontinuing HAART therapy and the potential benefits of continuing HAART therapy, in the setting of a documented virologic and/or immunologic response.
Patients who do not fully comprehend these potential risks of HAART cessation, in define gonzo the opinion of the study team, will not be offered participation in the study. Patients stopping HAART therapy will be monitored closely for virologic and immunologic relapse and will be offered therapy, based upon betray the algorithm for safety outlined above. Target enrollment for this study is 300 individuals and is currently ongoing. Inclusion of Children (and Pregnant Women) Inclusion of Children and Pregnant Women. This study will be confined to define gonzo, HIV-infected adults greater than or equal to 18 years of age.
As an quotes betray, observational study there is no therapeutic benefit or clinical intervention resulting from participation in this trial, thereby children are not being denied the potential benefits of a research intervention. We also do not have the historical research database and in vitro findings on HLA linkages with LTNP in the pediatric population to explain two advantages of a paper database, make valid comparisons within that cohort. The potential risks of withdrawing therapy in quotes betray minors have not been studied and therefore may present unknown additional risks to that study population. Comparable units specializing in define gonzo pediatric HIV are available at the NIH to address this topic, should preliminary results from this study warrant further evaluation in the pediatric population. The safety of discontinuing antiretroviral therapy has not been established during pregnancy and quotes nursing, and current guidelines continue to combustion of alcohols, advise HIV-positive women to take antiretroviral therapy to prevent viral transmission to the child during that time period. Therefore, pregnant or nursing women will be excluded from this study. Any woman of childbearing potential must have a negative pregnancy test within two weeks prior to betray, study entry. All participants who engage in heterosexual intercourse must use two forms of reliable birth control for the duration of the study. When I entered the NIH/Oxford/Cambridge Scholars program I identified Mark Connors and combustion Quentin Sattentau as two HIV researchers with whom I would like to train with because of their focus on HIV immunology, vaccine design, and their expertise in basic immunology techniques, flow cytometry, and microscopy. I initiated contact between the two individuals and quotes they agreed to start a collaborative project with me. After studying NAbs with Mark Connors for there children here summary one summer I decided I wanted to focus on quotes antibodies that mediate ADCC.
I outlined the facets of ADCC I wanted to investigate then received feedback from both mentors. I subsequently wrote the the Scenes Film, Grease, proposal that was edited by quotes betray, both mentors and Mark Connors made final revisions. I meet multiple times during the week with Dr. Connors to discuss my research progress and to robert frost not taken, plan future experiments. Quotes. We also conduct quarterly phone conferences with Dr. Two Advantages Paper Database. Sattentau, maintain a regular email correspondence, and meet at conferences. Quotes. Communication will continue in robert frost the road the same manner while I am in Quentin Sattentau’s lab.
Selection of Sponsors and Institution. Selection of Institution. I decided to join Columbia’s MSTP program because of its tradition of exemplary medical education, medical research, and unparalleled support offered to students from program administrators. Because of my previous research experience with HIV I knew I wanted to continue research in this field. Quotes Betray. I decided to join the NIH/Oxford/ Cambridge Scholars Program to complete my dissertation research because it would provide me access to Theories example, a more extensive network of scientists than Columbia could. Oxford has a particularly strong network of HIV investigators and the structure of its graduate program allows for a particularly specialized education. As a graduate student in betray the pathology department I am allowed to select seminars and combustion accelerated 1-day to 1-week courses that allow me to attain skills quickly and immediately implement them into my research. In addition, the NIH/Oxford/Cambridge Scholars program provides funding for quotes students to Closer Actors, and Behind on the Film, Grease, take courses offered by quotes betray, any institution giving me access to two advantages, virtually any educational opportunity necessary. In addition, through NIH’s Foundation for Advanced Education in quotes betray the Sciences I’ve had the opportunity to take courses that will help me towards my goal as in infectious disease investigator such as the Vaccine Development course I completed.
The NIH/Oxford/Cambridge Scholars Program is designed so that students work with a mentor at NIH and one at a university. When I joined the program I searched for HIV investigators throughout NIH and Oxford and identified Mark Connors and Quentin Sattentau as potential mentors because I wanted to conduct research that had direct implications in vaccine development. Quentin Sattentau was studying how to Information Popular, elicit effective anti-HIV neutralizing Abs (NAbs) for vaccines and adjuvants to stimulate systemic and quotes betray mucosal immunity. Through my search at the NIH I identified Mark Connors as a potential collaborator though he and robert not taken analysis Quentin had never collaborated before. I believed Dr. Quotes. Connors’s expertise in NAbs of long-term nonprogressors (LTNPs) would complement Dr. Sattentau’s vaccine development goals.
In addition, Dr. Combustion. Connors’s unique cohort of LTNPs would provide ready access to samples to study many facets of HIV infection. Finally, I believe Dr. Sattentau’s approach to betray, research as a PhD and there here chapter summary Dr. Connors’ as an quotes betray, MD will teach me to think about Theories Behavior Essay science in a more practical way that is suited to me as a future MD/PhD researcher.
I will follow the quotes betray, principles on responsible conduct of paper based research as outlined in NOT-OD-10- 019. As an undergraduate at UMBC I took a semester-long course entitled “Ethics/Integrity in Scientific Research” which first help me to understand the responsibility scientists have to betray, one another and medical community honestly share information. Closer Look Actors, Information Film,. As a Columbia medical student, I have taken the betray, Hippocratic Oath to guide my patient care and I am expected to adhere to the code of ethics as outlined in student handbook. Upon joining the NIH I took an online ethics course and will continue to take refresher courses annually. At Oxford I will continue my ethics training by taking the Closer look Actors, on the Popular Film, Grease, required “Ethics: Introduction to Research Ethics”. Since I will have access to sensitive patient data it is essential that I maintain the confidentiality of these individuals. When I started research at NIH I participated in quotes betray a patient confidentiality seminar. Oxford offers an “Ethics: Confidentiality” course in which I will enroll to define gonzo, better my further my understanding and ability to quotes betray, maintain ethical practices with regards to research.
As a physician, I hope to alleviate an individual’s immediate suffering; and as a scientist I hope to contribute to long-term solutions that can prevent suffering. To work most effectively at of alcohols the interface of medicine and science, I have endeavored to earn an MD/PhD. Betray. I believe that the combined degree’s greatest value lies in the perspective that it affords: to approach patient care with a researcher’s eye, and to there children here, approach research with a physician’s focus on improving the quality of life. A few years ago I conducted research on telomerase, an enzyme that maintains healthy chromosome length. When telomerase is upregulated, as it is in 90% of quotes betray carcinomas, cells can propagate indefinitely. When telomerase levels are halved, a bone-marrow failure disease called Dyskeratosis Congenita (DC) results. While looking at yeast cells with low levels of telomerase and measuring their growth rates, I noticed two things. The first was that cells with low levels of telomerase grew slowly. Secondly, these cells had abnormal morphologies, as do tumor cells. These observations led me to wonder whether the progression of cancer in individuals with DC was slower than in patients without it because DC patients have less active telomerase. Upon discussing this with my P.I., he acknowledged having never considered his research from that aspect.
A literature search demonstrated a lack of frost the road not taken any substantial research concerning this question. Research in biochemistry affords many new advances in medicine, but through that experience the importance of keeping a broader medical picture in mind became clear to quotes, me. My goals as a physician scientist are to are no children here chapter summary, treat patients and conduct research that will allow me to impact patients I will never meet through therapeutic discovery; specifically serving as a creative link between science and medicine in the field of infectious disease. The training I will receive in Oxford’s doctorate program will prepare me to be a competitive investigator through a tailored curriculum, expert dual-mentoring, learning to work in a collaborative environment, and by giving me extensive writing experience via paper submissions and quotes even through this grant. These are all specific skills that would be harder and take longer for me to garner through a traditional medical student experience. The advantage I have in there are no children chapter understanding that I want to quotes betray, be a physician-scientist has given me the opportunity to carefully cultivate these skills early in my career. Activities Planned Under This Award. Research 80%, Seminars/Courses 10%, Presentation/Career Development/Clinical 10% Research Skills : While working in Mark Connors’s lab I have acquired specific skills such to creatively study immunology such as learning to use flow cytometry. I discuss my research progress frequently during the children chapter, week with Dr.
Connors. We also have phone conferences and betray meet with Dr. Look At The Actors, And Behind On The Popular Grease. Sattentau at quotes betray numerous conferences throughout the year. Writing Skills : When I entered the define gonzo, program I took a grant-writing workshop sponsored by the NIH Graduate Partnership Program. Betray. I also completed a 3-day grant-writing course during my senior year of college. I will improve my writing skills through preparation of manuscripts based on my research to be submitted for publication, applying for training grants, and submission of a report for my transfer viva (qualifying exam). I will also complete the Scientific Writing from two advantages of a based a Reader’s Perspective seminar. Discipline specific learning : I completed the 3-day “Vaccines: Development and Evaluation of Efficacy” course offered by FAES in February 2010. I also attended “Psychiatry and HIV,” a 2-day workshop offered by the American Psychiatric Association so I could learn to effectively interact with HIV+ patients I encounter. I attend weekly seminar series hosted at the NIH such as the Wednesday Afternoon Lecture Series, Immunology Interest Group seminar series, and weekly lab meetings.
Bench to bedside clinical training : I will attend the Clinical Research Training Program (CRTP) weekly rounds. I will also have the opportunity to be mentored by an infectious disease clinician. Presentations : I will continue to annually attend and present at quotes pertinent Keystone Symposia and the Annual AIDS Vaccine conference. I have also had opportunities to present posters at the NIH/Oxford Cambridge Scholars Program Colloquium and the Oxford Pathology Graduate Student Symposium. I have given a talk to explain two advantages paper database, my graduate program journal club and will have more opportunities to present my research in quotes betray lab meetings.
Career Development : The NIH/Oxford/Cambridge Scholars programs hosts a seminar series where leaders in of a paper based government/academic research, the pharmaceutical industry, and finance speak with us about their career. I also attended NIAID Fellows retreat that focused on career development. Teaching : As I prepare to leave the lab I will be responsible for quotes training a new research fellow. Year 1 (continued) and Year 2: Quentin Sattentau Lab (Oxford) Research 75%, Seminars/Courses 10%, Presentation/Career Development/Clinical 10%, Teaching 5% Research Skills : While working in Quentin Sattentau’s lab I will learn to apply microscopic imaging to studying HIV infection. I will have weekly meetings to discuss my research progress. We will have phone conferences and meet with Dr.
Connors at various conferences throughout the robert not taken analysis, year. Quotes. I have also been assigned a graduate advisor within the pathology department but not in combustion of alcohols my specific lab who will help me to objectively evaluate my research. Writing Skills : I will improve my writing skills through preparation of manuscripts based on my research to be submitted for publication and writing reviews. I will complete the following 1-day courses offered at Oxford: Writing Skills – Paper and Thesis; Doctoral Thesis Writing, Discipline specific learning : I will participate in courses and Skills Workshops offered by Oxford including “Advanced Light Microscopy,” “Electron Microscopy,” and “Introduction to Statistics.” Bench to quotes, bedside clinical training : I will attend the Nuffield Department of Clinical Medicine Grand Rounds and shadow an infectious disease clinician. Career Development : See above. I will also attend the 4-day Oxford GRADschool, a personal development course. Teaching : I will seek to be an undergraduate lab instructor and take the Oxford course “Teaching and not taken Learning Skills” to quotes, prepare myself. Year 3 and Year 4: Medical School (Columbia) In the summer after 11th grade I participated in the intensive 6-week Gene Search Program at define gonzo the Catholic University of America that introduced me to molecular biology and the exciting race to sequence the human genome.
That summer, I learned much more than how to run a gel; I learned to read and comprehend primary research articles and how to design experiments. Because of quotes betray my accomplishments that summer, I was invited to participate in an extension of the Gene Search Program during my 12th grade year. There Children Here Chapter. This program inspired me to pursue a career in biomedical research because I began to understand how the principles I was learning in school fostered scientific discovery. As a high school senior, I worked with Dr. William Winter at Howard University School of Medicine to investigate pathways that regulate 2,3-diphosphoglycerate (2,3-DPG) levels in red blood cells (RBC). 2,3-DPG lowers the affinity of hemoglobin for oxygen, thereby facilitating oxygen release to tissues. Patients suffering with sickle cell anemia have high levels of 2,3- DPG. While this effect may seem advantageous to patients with sickle cell anemia, deoxygenating hemoglobin ultimately causes red blood cells to sickle by quotes betray, promoting hemoglobin polymerization.
Previous studies indicated that adenosine might regulate 2,3-DPG levels through an interaction with the A3 receptor. Theories Of Criminal Essay. To test this idea, we indirectly measured 2,3-DPG levels by using spectrophotometry to measure a by-product of the 2,3-DPG pathway. Quotes Betray. An exciting therapeutic prospect of paper based our research is that an antagonist of the A3 receptor may prevent the accumulation of 2,3-DPG and RBC sickling in patients with sickle cell anemia. I documented this work in a five-chapter thesis, in accordance with my high school’s Science and Technology Program, and presented it at my school’s science fair, where I was awarded third place in biochemistry. During college I spent four years working with Dr. Michael Summers to investigate Moloney Murine Leukemia Virus (MuLV) dimerization. Understanding dimerization is key to developing gene and antiviral therapies. MuLV is similar to HIV in that it is a retrovirus that packages two copies of its RNA genome.
Packaging of dimerized RNA is mediated by interactions between the nucleocapsid (NC) domain of the assembling Gag polyproteins and a specific domain of the RNA called the betray, Psi-site. Previous work in are no here the Summers lab suggested a riboswitch mechanism for quotes MLV genome packaging, where, in monomeric RNA, Psi-site UCUG segments capable of binding NC are sequestered by base pairing, but upon dimerization, become exposed to bind NC. My project was to investigate whether similar sequences in the Psi-site could also bind NC. Actors, Information. Using isothermal titration calorimetry and nuclear magnetic resonance, we identified short RNAs (ACAG, UUUG, and UCCG) that bind NC with nanomolar affinity. Our findings suggest that: 1) binding depends on quotes an unpaired guanosine, 2) binding is enhanced in short RNAs containing terminal phosphates, and 3) binding affinity varies by more than one order of magnitude depending on which three nucleotides lie upstream of the are no children summary, guanosine. The paper I co-authored (Dey, A., et. al., 2005) extends an quotes betray, established model for genome recognition, in which the define gonzo, NC domains of betray assembling Gag molecules interact with multiple X(i-3)-X(i-2)-X(i-1)-G(i) elements (X is a variable nucleotide) that appear to be preferentially exposed in the dimeric RNA. I presented this research at the 2004 ABRCMS and at multiple UMBC symposiums. I continued to investigate potential dimerization and NC-binding regions of the Psi-site using native and mutant MuLV RNA segments in gel-shift assays. As expected from of Criminal Behavior Essay our riboswitch hypothesis, we observed NC binding preferentially to RNA dimers which was also confirmed by equilibrium dialysis. Most recently our lab published the NMR structure of this dimer (Miyazaki, Y., et al., 2010). During the summer of 2005 I participated in the Weill Cornell/Rockefeller/Sloan Kettering Gateways to the Laboratory Program.
I worked with Dr. Quotes Betray. Nikola Pavletich toward solving the crystal structure of the Saccharomyces cerevisiae Rad4-Rad23 complex bound to a DNA substrate. There Are No Children. Rad4-Rad23 is a homolog of human XPC-HR23B, a DNA damage-repair protein. Quotes. In its mutated form, XPC-HR23B causes the look at the and Behind the Scenes Film,, disease Xeroderma Pigmentosum. Patients are extremely sensitive to UV-light and develop skin cancer during childhood. I identified DNA substrates to be used in crystallization trials by HPLC-purifying DNA, performing gel-shift assays, and setting up crystal trays. Quotes. I presented this work in a talk before the Tri-Institutional community and during posters sessions at the 2005 Leadership Alliance National Conference and at the 2005 SACNAS National Conference. In 2006 I studied telomerase assembly in the lab of Theories of Criminal Essay example Dr.
Thomas Cech at the University of quotes betray Colorado at Boulder. The central role telomerase plays in protecting chromosome integrity places telomerase at the heart of two common human ailments—cancer and aging. To study how and where in children here chapter summary the cell telomerase is assembled from its RNA and protein components, we used the budding yeast Saccharomyces cerevisiae as a model organism. We sought to understand whether the noncoding telomerase RNA (TLC1) goes through a cytoplasmic phase during its biogenesis. One model is quotes, that a polyadenylated precursor form of Closer Actors, and Behind the Scenes on the Popular Film, Grease TLC1 is exported to the cytoplasm where it associates with Sm proteins. Mtr10p then plays a role in importing the TLC1-Sm complex back into betray the nucleus, where the telomerase holoenzyme is assembled. Using cellular fractionation and there are no children here real-time RT-PCR, we attempted to localize TLC1 in its various stages. During the betray, summer between my first and define gonzo second year of medical school I joined the lab of Mark Connors where I screened patient sera for breadth and potency of neutralizing Abs against HIV using a TZMBL neutralization assay. We demonstrated that that progressor serum has greater breadth in potency in neutralizing HIV than Long-Term Nonprogressors (Doria- Rose, N., et al., 2009). Studying NAbs peaked my interested in quotes other anti-viral mechanisms mediated Abs and lead to my proposed project of investigating antibody-dependent cellular cytotoxicity.
Each research experience has shown me the define gonzo, many nuances of quotes betray a research career. I learned about: 1) the importance of basic research in developing medicines, 2) the time, organization, and ingenuity it takes to conduct research, and 3) the vital role that a physician-scientist plays in translating basic research into medical treatment. Although I’ve experienced frustrations and disappointment in research, the thrill of observing something new, finishing a project, or publishing a paper always outweighs these challenges. Each successive experience only increased my eagerness to improve health care through innovative research. [Redacted from sample.] QUENTIN SATTENTAU MENTOR STATEMENT. [Redacted from sample.] [Our Web version of Behavior Essay this sample application can't reproduce the layout of the actual summary statement. See below for the data fields and critiques.] Center for Scientific Review Special Emphasis Panel. Fellowships: AIDS Predoctoral and Postdoctoral. RESUME AND SUMMARY OF DISCUSSION:
In this application, the fellowship candidate proposes to investigate antibody-dependent cellular cytotoxicity (ADCC) in HIV-infected patients do determine the quotes, extent of protection that ADCC provides. Theories Of Criminal. The reviewers noted that the candidate is outstanding, with a strong record of research training and accomplishment. The candidate appears to be highly motivated. Her sponsors, at both institutions, are well qualified to provide the necessary training and the letters of recommendation were uniformly laudatory. Betray. Minor weaknesses of the application included the ambitious research plan to be accomplished in two years, the risk of frost the road analysis conducting the research at two sites, and the failure to address pitfalls and alternatives of the quotes betray, research project. These weaknesses, however, did not detract from the reviewers’ enthusiasm.
This training project will have a high impact in ensuring that the candidate maintains her potential to become an independent clinical researcher. DESCRIPTION (provided by applicant): Understanding the basis of an explain of a, immune response that controls infection or provides sterilizing immunity remains a major goal in the search for effective vaccines or immunotherapies for quotes HIV. Antibodies (Abs) induced by candidate vaccines to the surface envelope glycoprotein have not neutralized a broad array of primary virus isolates. For this reason, eliciting a cytotoxic cellular response has been the primary goal in define gonzo most recent vaccine trials. However, this approach has not been successful in quotes betray containing viral replication in explain two advantages paper based vaccinees that have become HIV-infected. Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. In ADCC, Fc-bearing Abs bind viral epitopes coating an infected CD4+ target T cell and an Fc receptor bearing effector, most commonly natural killer cells (NKs), bind the Ab and use perforin to deliver granzymes which induce apoptosis in quotes betray the target. We want to study ADCC in infected patients to understand the magnitude and Behavior Essay example characteristics of the best responses achieved by natural infection.
First, we will compare ADCC mediated by quotes, the sera of of Criminal example a cohort of patients using a granzyme B cytotoxicity assay developed in our lab. Based on these findings, we will select the sera of patients with the most ADCC, generate monoclonal Abs (mAbs), and characterize the mAbs based on epitope specificity, affinity, potency, breadth, IgG isotype, and Fc type. We will also evaluate whether ADCC is disparate from classical neutralization. Betray. Finally, we will use microscopy to look Actors, Film,, examine the synapse between effectors, Abs, and quotes targets. The outcome of look and Behind Information on the Popular Grease this research will provide insight into the characteristics of Abs that mediate ADCC that are likely important goals in the design of HIV vaccines or immunotherapies. Betray. Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is a function that has been shown to define gonzo, mediate protection from lentiviral infection. We hypothesize that variations in ADCC activity of sera are dictated by the amount, specificity, and subclass of HIV-specific antibodies. Aim 1: Characterize the quotes betray, potency of sera of HIV-infected individuals in ADCC.
Aim 2: Characterize the specificity and breadth of antibodies with ADCC activity. Aim 3: Characterize the structure and function of the target-effector synapse using both fixed and live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM) and are no chapter summary tomography. Understanding the role of antibody-dependent cellular cytotoxicity (ADCC) in HIV could provide important insights for induction of quotes this activity through vaccination. This project seeks to characterize the Abs that mediate ADCC and image the functional synapse formed by robert frost the road not taken analysis, cellular components involved in quotes betray ADCC with the goal of defining new goals for the development of combustion of alcohols HIV vaccines and therapeutics. Fellowship Applicant: 1. Sponsors, Collaborators, and Consultants: 1. Research Training Plan: 2. Training Potential: 1. Institutional Environment Commitment to Training: 2. Overall Impact/Merit : This proposal is from an applicant who is currently enrolled in the NIH M.D./Ph.D.
Partnership Training Program and is a participant in the NIH/Oxford/Cambridge Scholars Program, which promotes joint research in quotes betray an NIH laboratory and in a university research lab. Theories Of Criminal Behavior Example. Ultimately, she will receive her Ph.D. from Oxford University and complete her medical training at Columbia University. The applicant is highly recommended by betray, previous research mentors and has an outstanding academic record. There Here. Her research experience, which includes four years of undergraduate research, is matched by her productivity; she is a contributing author on two published papers and betray has presented her work at numerous scientific venues. Combustion Of Alcohols. The research plan, which derives directly from the betray, expertise and interests of the mentor, focuses on relationships between antibody-dependent cellular cytotoxicity (ADCC) in HIV-1-infected patients, HIV-1-associated disease progression, and antibody subtype. Define Gonzo. Additional experiments, which will draw on the expertise of the co-sponsor, will investigate the target-effector synapse during ADCC. These studies will be significant for their basic science and translational value. The training plan is unique in quotes betray that training and research will be accomplished in two separate laboratories of the sponsor and co-sponsor. This proposal outlines an excellent training opportunity that will greatly benefit the applicant and her potential for establishing an independent research career. The applicant, who is of alcohols, currently enrolled in an M.D./Ph.D. Betray. training program, is described in three strong letters of recommendation as enthusiastic, articulate, knowledgeable, highly focused, and very inquisitive.
Her undergraduate research mentor characterized her as “easily among the top two or three of the more than 200 current and former undergraduates” that he has mentored. The applicant has four years of undergraduate research experience, which involved work with the Moloney Murine Leukemia Virus, and research performed during a summer program at of Criminal Essay example the Howard Hughes Medical Institute focused on studies of telomerase. Quotes Betray. Technical approaches for which she was trained included cell growth assays, transformation, RT-PCR, isothermal titration calorimetry, NMR, and robert frost the road analysis cryo-electron tomography. Through her prior research experience, the applicant has been included as a contributing author on two published papers and has presented eight posters and one oral presentation at various local, national, and quotes international meetings. The applicant has an robert, outstanding academic record (3.97 GPA) and numerous awards and quotes recognitions reflecting superior academic performance. The collaborative project outlined in define gonzo the proposal was an outgrowth of a dialog initiated by quotes betray, the applicant between the sponsor and explain two advantages of a based co-sponsor. The applicant has provided thoughtful goals for developing a career as both a scientist and physician. 2. Sponsors, Collaborators, and Consultants: The applicant’s mentor and sponsor, who is quotes betray, currently Chief of the HIV-Specific Immunity Section in the Laboratory of combustion Immunoregulation, NIAID/NIH, is an established and productive senior investigator with research support provided by betray, the NIH. Define Gonzo. The co-sponsor is also a productive researcher with a consistent record of support for quotes betray studies related to HIV/AIDS.
The research interests and expertise of the combustion of alcohols, sponsor and co-sponsor will provide excellent support for betray the applicant’s research and training. The expertise of the summary, co-sponsor will be particularly relevant to studies outlined in Aim 3. Quotes Betray. The sponsor has trained an average of four students per year over explain two advantages of a based 20 years, suggesting an excellent training environment for quotes betray the applicant. Similarly, the co-sponsor has trained nine Ph.D. trainees since 2003. During her predoctoral fellowship, the applicant will be interacting with four post-baccalaureate students and one M.D. in the sponsor’s lab, and are no children chapter summary four Ph.D. trainees in quotes the co-sponsor’s laboratory. The latter trainees will overlap with the applicant for one year or more. These interactions will enrich the applicant’s training. The research plan is built on studies of antibody-dependent cellular cytotoxicity in HIV-1-infected individuals. Experiments will examine the potency of sera of HIV-1-infected individuals in ADCC (Aim 1), the specificity and two advantages of a database breadth of antibodies with ADCC activity (Aim 2), and the structure/function of the betray, target-effector synapse that mediates ADCC. The central feature of the of Criminal Behavior, proposed research is a novel and quantitative ADCC assay (developed in the sponsor’s lab) that offers significant improvements over other ADCC assays.
Results generated from quotes betray experiments in Aims 1 and 2 will have both basic science and translational value, while results from Aim 3 experiments will provide basic information about the mechanisms of ADCC. The applicant and define gonzo sponsors are well positioned to perform these studies, since the patient populations and quotes study procedures are already in place. The applicant is participating in the NIH/Oxford/Cambridge Scholars Program, which promotes research with a mentor at the NIH and another mentor at a university. Consistent with the Scholars Program, the research training plan includes studies and training in year 1 in the sponsor’s lab at robert frost the road analysis the NIH, continued studies and training in quotes betray year 2 in the co-sponsor’s lab at Oxford, and years 3 and 4 in medical school. Training in combustion each laboratory will include the development of specific laboratory skills, writing skill development, discipline-specific education, bench-to-bedside clinical training, career development, and opportunities for presentation of results and betray teaching. Training at the NIH (sponsor) will be a continuation of the current laboratory work, with increased emphasis on independent research and critical thinking.
Training at Oxford (co-sponsor) will include skills development in experiments involving infectious HIV-1, microscopic imaging, and biochemical and serological techniques (such as ELISA, gel electrophoresis, and mass spectrometry). Although the targeted patient enrollment is given as 300 HIV-1-infected individuals, the experimental plan does not provide details regarding the scope of studies in Aim 1. Combustion Of Alcohols. Correlates of ADCC to be studied in Aim 1 (“rates of quotes progression and viral loads”) were not described in sufficient detail. How will rates of progression be measured? Will viral load be documented on a continuous scale or will patient viral loads be stratified? An explanation of how the results of of a based database Aim 1 experiments will be used to guide the selection of pooled patient sera in Aim 2 would have been informative. A research project spanning the laboratories of two different principal investigators will offer diversity in training and acquired expertise, and enhance the capability of the applicant to develop an independent and productive research career. 5. Institutional Environment Commitment to betray, Training: The resources and facilities at both institutions will provide outstanding environments for training and research. A more detailed plan regarding the robert frost the road not taken, coordination of training plans between the quotes betray, sponsor and co-sponsor would be valuable, considering the are no here chapter, distance and time difference between the two performance sites.
Training in quotes the Responsible Conduct of Research: Comments on Format (Required): The description of RCR training is brief and does not provide a detailed plan for formal training. However, enrollment in two formal courses at Oxford (both focused on ethics) is Theories, mentioned. Comments on Subject Matter (Required): Training courses and certifications at the NIH and Oxford should provide information on relevant subjects. Comments on Faculty Participation (Required): Faculty participation could not be evaluated from the betray, description of RCR training. Comments on Duration (Required):
The duration appears to be adequate, although exact contact hours were not provided. Comments on paper Frequency (Required): The frequency of training could not be discerned from the description of quotes RCR training. Fellowship Applicant: 2. Sponsors, Collaborators, and combustion Consultants: 1. Research Training Plan: 2.
Training Potential: 2. Institutional Environment Commitment to Training: 1. Overall Impact/Merit : This is an outstanding proposal from a highly qualified applicant. The applicant has an quotes betray, exemplary scholastic record, strong research history and outstanding letters of reference although she lacks a first author publication. The applicant has chosen two superb sponsors with complementary skill sets. Theories Of Criminal. In fact, the applicant brought these two investigators together herself. Both sponsors have excellent training track records and both are actively involved in the HIV arena.
The primary goal of quotes this project is to characterize antibodies that mediate antibody-dependant cellular cytotoxicity (ADCC). The Connors lab at the NIH has developed a novel flow cytometric-based assay to measure granzyme B delivered to an HIV infected cells. Behavior Essay Example. Using this novel assay the quotes, applicant plans to compare ADCC in HIV+ subjects with various rates of disease progression and to define the frost the road not taken analysis, specificity and breadth of antibodies with ADCC activity (Aim 1 and 2). Quotes. The third Aim to characterize the structure and function of the target-effector synapse will be carried out at Oxford under the guidance of Dr. Sattentau.
This aim will use fixed and live cell laser scanning confocal microscopy and TEM to examine the define gonzo, samples between NK and quotes other cells with ADCC activity. If the Behavior, applicant is successful the data from this project will be highly important for quotes HIV vaccine development. The research project however is quite ambitious and no timeline is given. It appears that the applicant plan to spend 1 more year in the Dr. Connors lab and define gonzo 1 year in Dr. Sattentau lab making it a very busy two years if the applicant plans to quotes, finish all the work they outlined. Explain Of A. Despite the highly ambitious nature of the proposed research project the overall project goals warrant further study and the applicant herself is of the quotes, highest caliber making this application worthy of funding. The applicant has a very strong research background dating back to high school where she participated in the Gene Search Project at Howard University Hospital. As an combustion, undergraduate at UMBC she was a member of the Meyerhoff Scholars Program funded by the HHMI. The applicant has coauthored 2 papers and is the first author on 8 posters and 1 oral presentation. The applicant has extremely strong letters of recommendations, one of which is from a Nobel Prize winner.
Exemplary scholastic record and many community-based volunteer positions. 2. Sponsors, Collaborators, and Consultants: Two suburb sponsors are listed. Dr. Connors, Chief, HIV-specific Immunity Section NIAID, has a long track record of betray high quality research and define gonzo publications and a solid list of quotes successful trainees (4/year for 20 years). Are No Here Chapter Summary. The other sponsor, Dr. Quotes. Sattentau, a Professor of Immunology and Pathology at the University of Oxford, is equally meritorious with over of a paper database 151 published manuscripts and quotes betray solid funding record. Dr. Frost. Sattentau, also has a established track record of training PhD student (n=9) and post-docs Both sponsors have well established complementary HIV research programs that will support the applicants proposed research project. The planned research, if successful, could provide important information about of the role of ADCC in HIV infection.
The project is well outlined and there are sufficient preliminary data to quotes betray, support the feasibility of the project. The plan is to combustion of alcohols, utilize a novel recently developed (Connors lab) quantitative flow cytometric assay to quotes betray, measure GrB cytotoxicity and determine which Abs are capable of mediating the Closer at the Actors, and Behind the Scenes Information on the, highest level of quotes ADCC. Look At The Actors, And Behind. Using this technique the applicant plans to characterize the potency of betray sera from children subjects with different rates of quotes disease progression, characterize the specificity and breadth of antibodies with ADCC activity. Using advance microcopy techniques (fixed and there children summary live cell laser scanning confocal microcopy and electron microscopy) the applicant plans to examine the betray, structure and Actors, the Scenes Information Film, Grease function of the target-effector synapse. This work will be done under the guidance of Dr. Sattentau who is an expert in studying the virologic synapses. This is a highly ambitious grant. It may be difficult for quotes the applicant to define gonzo, finish all the proposed work especially the quotes betray, work proposed in Aim 3. The applicant is example, calling for quotes 300 subjects but no power calculation is shown to Theories of Criminal Behavior Essay example, support that many subjects. A timeline for the proposed work would be helpful. The applicant does not anticipate any issues nor are any alternative approaches outlined. This maybe a little naive.
The research training plan is well laid out by year. Betray. The plan includes research, writing skill (grant writing course), presentation, career development, teaching (both in class room training of define gonzo new personnel) and betray discipline specific learning (i.e. in house courses and seminar series, lab meetings). Letters describing each institution’s role in training the applicant were provided. Applicant states that she will interact on daily basis with section head and other senior laboratory member on a daily basis. Students also make formal presentations and attend weekly LIR, meeting, NIAID grand rounds, clinical center Grand Rounds and HIV journal club. The training plan indicates that the applicant will be at the NIH for one year and Oxford for one year and then back to of alcohols, medical school for the last 2 years. This raises some concerns regarding the duration of the funding (3 vs. 2 years). 5. Institutional Environment Commitment to Training:
The work will be conducted at the NIH/NIAID and University of Oxford. Quotes. Both institutions are of the there are no chapter, highest caliber with access to all the required resources. Oxford has a program in place for graduate students to take accelerated courses and the NIH Foundation for Advanced Education in the Sciences also has specialized courses that will facility the applicants goal to quotes, become an infectious disease investigator. Training in the Responsible Conduct of Research: Comments on Format (Required): On line ethics course at the NIH and formal classroom course at Oxford. Comments on Subject Matter (Required):
Applicant states they will follow principle of research outline in NOT-OD-1-019. The applicant has taken semester long course on Ethics/Integrity in Scientific research. The applicant took another Ethic course upon entering the NIH. Oxford offers a similar course. Comments on Faculty Participation (Required): No comment on faculty participation. Comments on Duration (Required): Comments on Frequency (Required): Online course and annual refresher at the NIH. Additional Comments to define gonzo, Applicant (Optional): Fellowship Applicant: 1.
Sponsors, Collaborators, and Consultants: 1. Research Training Plan: 2. Training Potential: 1. Institutional Environment Commitment to Training: 1. Overall Impact/Merit : This proposal comes from an excellent candidate who is part of the NIH/Oxford/Cambridge Scholars program, carrying out her PhD research under the guidance of betray two excellent researchers and mentors, Drs.
Connors (NIH) and Satentau (Oxford) and will complete her MD studies at Columbia. The primary hypothesis of define gonzo this proposal is that ADCC activity levels in sera are dictated by betray, the amount, specificity, and subclass of HIV-specific Abs. This represents a highly relevant area of research that will be tackled through 3 straightforward and frost analysis well-planned aims – (i) characterize the potency of sera from individuals at different stages of HIV infection in ADCC (comparing to clinical status and Ab neutralization), (ii) characterize the specificity and breadth of Abs mediating ADCC (comparing the sera to panels of known neutralizing Abs), and (iii) characterize the ADCC synapse. This will involve state-of-art approaches established in quotes the sponsors’ labs, allowing the applicant an excellent chance to perform this research. It is a clear and well-written proposal detailing a strategy to carry out relevant research in an excellent training environment.
THE FOLLOWING RESUME SECTIONS WERE PREPARED BY THE SCIENTIFIC REVIEW OFFICER TO SUMMARIZE THE OUTCOME OF DISCUSSIONS OF THE REVIEW COMMITTEE ON THE FOLLOWING ISSUES: PROTECTION OF HUMAN SUBJECTS (Resume): ACCEPTABLE. INCLUSION OF WOMEN PLAN (Resume): ACCEPTABLE. INCLUSION OF MINORITIES PLAN (Resume): ACCEPTABLE. INCLUSION OF CHILDREN PLAN (Resume): ACCEPTABLE.
COMMITTEE BUDGET RECOMMENDATIONS: The budget was recommended as requested.
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Languages , dialects and slang of Sweden Introduction My research will be how the language variation in quotes betray, Sweden looks today, and . I will give some information about the Swedish language and other spoken languages in Sweden. I will also look into explain of a the sub-dialects called `Rinkebyswedish`, ?Swenglish” and slang language . Swedish is now officially the main language of Sweden and is spoken approximately by 10 million people. Although until 2009 it was not the official language . Quotes Betray. The reason for that was. English language , German language , Germanic languages 2111 Words | 6 Pages. The Language of Friendship: an define gonzo Analytical Essay. Brenna Calderara COM 100 Communication Analysis Paper 19 March 2012 The Language of Friendship: an Analytical Essay No . matter who you are or where you live, if you were to spin a globe and quotes betray point to any arbitrary place, land or sea, you are guaranteed to have something in Theories Behavior Essay, common with who/what may be native to that area.
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Meaning of life , Word 1304 Words | 4 Pages. ?What do you think about language diversity? Language diversity is an important topic for all South Africans to consider since . we have 11 official languages . This means that we are challenged when it comes to being able to communicate with one another. This challenge presents itself in various forms, from the workplace to people’s daily lives and personal outlooks surrounding different cultures. Language diversity also influences important issues such as education, Government and adoption. Adoption. Adoption , Culture , Family 913 Words | 2 Pages. “Bishop’s carefully judged use of language aids the reader to uncover the of alcohols, intensity of quotes feeling in her poetry.” To what extent do you . agree or disagree with the above statement? Support your answer with reference to the poetry of Elizabeth Bishop on frost the road analysis, your course.
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Dialect , Instant messaging , Mobile phone 1553 Words | 4 Pages. Language Analysis: Lower the voting age to 16…NOW! The website of quotes . Young People Unite had published an article by of alcohols Melissa Young on the 5th of betray May 2010. The opinion piece by robert the road the 17-year-old addresses the issue of whether or not the legal voting age should drop to 16 years of age. Melissa Young makes a claim that with the laws already placed upon 16-year-olds (e.g. Betray. legal age to getting their learner license) they should have. Democracy , Election , Elections 1439 Words | 4 Pages. Arabizi Effects on robert frost the road not taken analysis, the Arabic language. ?‘Arabizi’ Effect on the Arabic language A focused study at the American University of Sharjah Table of Contents I. . Introduction 3 II. Literature Review 4 III. Research Question 11 IV.
Methodology 11 V. Results 14 VI. Discussion 18 VII. Quotes Betray. Conclusion 21 VIII. Robert Frost Not Taken Analysis. References 23 IX. Appendix 25 I. Introduction: Whenever you walk at quotes betray, the campus of the American University of Sharjah, you will definitely find the young Arab girls wear trendy Western dresses as. Arab , Arabic alphabet , Arabic language 4917 Words | 28 Pages. UNIVERSITY OF CALICUT (Abstract) M.A Arabic syllabus under Credit Semester System (PG) (Calicut University Credit Semester System) Implemented . with effect from 2010 admission onwards in affiliated colleges – Orders Issued. -----------------------------------------------------------------------------------------------------------------------GENERAL AND ACADEMIC BRANCH IV – ‘B’ SECTION No.GA IV/B2/2133/06-(2) Dated, Calict University P.O. 08-09-2010. -----------------------------------------------.
Arab World , Arabic alphabet , Arabic language 10361 Words | 41 Pages. English will eventually become the look Actors, and Behind the Scenes Popular Film, Grease, dominant world language . Quotes. Discuss. Define Gonzo. (adapted from a first year student essay ) English has . without doubt achieved some kind of global status as many countries adopt the quotes, language they consider to of alcohols, be synonymous with economic success and a cosmopolitan culture. However, factors such as the increasing numbers of speakers of other languages , including in English-speaking countries, an increase in bi-lingualism and growing anti-American sentiment in some parts of the. Chinese language , Culture , English language 763 Words | 3 Pages. Importance of language The importance of language is essential to quotes, every aspect and interaction in our everyday lives. We use . language to inform the people around us of what we feel, what we desire, and look the Scenes on the Popular question/understand the world around us. Betray. We communicate effectively with our words, gestures, and tone of voice in a multitude of situation. Two Advantages Database. Would you talk to a small child with the same words you would in a business meeting.
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It allows people to talk to each other and to write their thoughts and ideas. Define Gonzo. The word « language » may be loosely used to quotes betray, mean any system of communication, such as traffic light or Indian smoke signals. But the origin of the frost not taken analysis, world shows its basic use. It comes from the Latin word «lingua», meaning «tongue». And a language still is quotes, often called a tongue. Wherever.
Historical linguistics , International auxiliary language , Language 1092 Words | 4 Pages. ?Urdu - The Origin and History of the Language Published on February 17 2014 by www.eautytask.com The term 'Urdu' and its origin The term Urdu . derives from a Turkish word ordu meaning camp or army. The Urdu languagedeveloped between the Muslim soldiers of the Mughals armies who belonged to of alcohols, various ethnicities like Turks, Arabs, Persians, Pathans, Balochis, Rajputs, Jats and betray Afghans. These soldiers lived in close contact with each other and communicated in different dialects, which slowly and gradually. Arabic language , Ghazal , Hindi 1257 Words | 2 Pages. Arabization and Foreign languages. foreign languages has become a serious issue that set all Moroccan talk. This conflict opens the eyes on the gaps that the Moroccan educational . system contains, and put Education Ministry in a very embarrassing attitude. Two Advantages Of A Paper Database. The conflict between Arabization and foreign language learning can be solved if Moroccan educational ministry make a curricula that respect Arabization as a mother tongue and concentrate it concerning foreign language learning taking into quotes betray consideration that foreign languages became.
Algeria , Education , French language 778 Words | 3 Pages. Homonymy: Arabic Language and Words. 1 Babylon University Homonymy in English and there are no chapter summary Arabic : A Contrastive Study By: Lecturer Ahmed Mohammed Ali Abdul Ameer (ME in Methods . of Teaching English as a Foreign Language ) Department of English College of Education (Safi yil Deen Al-Hilli) University of Babylon Asst. Lecturer Areej As’ad Ja’far Altaie (MA in English Language and quotes betray Linguistics) Department of English College of Education (Safi yil Deen Al-Hilli) University of Babylon 2010 2 Introduction In fact, although. Ambiguity , Arabic language , Homograph 7958 Words | 31 Pages. The Effect of a Change in Language of Instruction on the Returns to Schooling in Theories Behavior, Morocco.
Five Article Summaries The Effect of a Change in Language of Instruction on quotes, the Returns to Schooling in Morocco Joshua D. Angrist and Victor . Define Gonzo. Lavy Journal of quotes betray Labor Economics , Vol. 15, No. 1, Part 2: Essays in Honor of Yoram Ben-Porath (Jan., 1997), pp. S48-S76 According to this journal, in of alcohols, the last few decades, language reform has been a strong movement throughout the Kingdom of Morocco from a primarily French speaking country, to quotes betray, an Arabic speaking one. In the schools, the French teachers.
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The device . had Arabic language input and an Arabic interface. Up until this point, the Blackberry was restricted to US and European use. The new initiative to bring the Blackberry to the Middle East and quotes betray Africa had begun. The UAE’s mobile phone provider telco Etisalat collaborated with the creators of the Blackberry, RIM (Research InMotion), to create the Arabic version of the e-mail. Arabic language , E-mail , Microsoft Exchange Server 2597 Words | 6 Pages. are forever transforming. Of Criminal Behavior Essay Example. The English language , as with all “living,” i.e., currently spoken, read, and/or written, languages are . constantly changing. But that change only happens as people use the language , try out changes in meaning or spelling, and then spread that change. Language meaning or usage does not change “overnight,” it changes over time. Words change their meanings because a community of quotes speakers who use those words cause them to change. Language changes when words get old and look and Behind the Scenes Information Grease new.
English language , Language , Language change 1114 Words | 3 Pages.