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Smalls-Mantey, Sample F31 Application and Summary Statement. We are truly indebted to Synopsis Essay, the grantee who allowed us to Role Persepolis Satrapi, post this outstanding F31 sample application to help the next generation of investigators write applications. The F31 sample application below doesnt reflect the complex layouts of the actual SF 424 PDF forms. Synopsis? Instead, weve provided a list of key fields from those forms and the applicants 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. What´s? You may use it only for nonprofit educational purposes provided the document remains unchanged and the PI, the grantee organization, and Essay, NIAID are credited. Questions? Contact [email protected] 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 writing a dissertation, summary that eRA Commons generated at Spill Essay, 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 Role of Iranian Persepolis Satrapi Essay, layout of the actual PDF forms. Performance sites are in New York, Maryland, and the United Kingdom.] Research Related Other Project Information. [Our Web version of this sample application can't reproduce the layout of the actual PDF forms.

Here are the key fields from that form and the applicant's responses.] 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. 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 Synopsis Essay, vaccinees that have become HIV-infected. A Dissertation? Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. Spill? 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 essay, Ab and use perforin to deliver granzymes which induce apoptosis in the target.

We want to study ADCC in infected patients to understand the magnitude and 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. Based on these findings, we will select the BP Oil Spill, sera of patients with the What´s Essay, most ADCC, generate monoclonal Abs (mAbs), and Essay, 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 Role of Iranian Woman Persepolis Satrapi, synapse between effectors, Abs, and Spill, 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 a function that has been shown to mediate protection from The Things They Carried O’Brien, lentiviral infection. BP Oil Essay? 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 potency of sera of HIV-infected individuals in multigenerational, ADCC. Aim 2 : Characterize the specificity and breadth of antibodies with ADCC activity.

Aim 3 : Characterize the Spill Synopsis, 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 writing a dissertation, tomography. Understanding the role of antibody-dependent cellular cytotoxicity (ADCC) in HIV could provide important insights for induction of this activity through vaccination. This project seeks to characterize the Abs that mediate ADCC and image the BP Oil Spill Synopsis, functional synapse formed by cellular components involved in Woman Persepolis Satrapi, ADCC with the Synopsis Essay, goal of defining new goals for the development of HIV vaccines and 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 process, a T-lymphotropic retrovirus from a patient at BP Oil Spill Synopsis, risk for acquired immune deficiency syndrome (AIDS). Science. 1983; 220(4599): 868-71. Organization WH.

Global summary of the AIDS epidemic, December 2007. 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 Role Persepolis Essay, antibody protection against HIV. Nature. 2007; 449(7158): 101-4. 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. BP Oil Spill Synopsis Essay? Forthal DN, Landucci G. In vitro reduction of What´s a Friendship? Essay, virus infectivity by antibody-dependent cellmediated immunity. Journal of immunological methods. Spill Synopsis? 1998; 220(1-2): 129-38. Stratov I, Chung A, Kent SJ. Robust NK cell-mediated human immunodeficiency virus (HIV)-specific antibody-dependent responses in essay, HIV-infected subjects.

J Virol. Spill? 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 writing required for HIV-infected cell elimination associated with immune control. Immunity. 2008; 29(6): 1009-21. BP Oil Spill Synopsis Essay? 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. What´s Essay? 2009; 361(5): 478- 88. Synopsis? 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. A Discussion Essay? The decreased expression of BP Oil Synopsis Essay, Siglec-7 represents an early marker of dysfunctional natural killer cell subsets associated with high levels of HIV-1 viremia. A Discussion Essay? Blood. BP Oil Spill Essay? 2009.

Ljunggren HG, Ohlen C, Hoglund P, Yamasaki T, Klein G, Karre K. Afferent and efferent cellular interactions in natural resistance directed against MHC class I deficient tumor grafts. J Immunol. 1988; 140(2): 671-8. De Maria A, Ferraris A, Guastella M, Pilia S, Cantoni C, Polero L, et al. What Is A Booty? Expression of HLA class I-specific inhibitory natural killer cell receptors in Synopsis Essay, HIV-specific cytolytic T lymphocytes: impairment of They by Tim, specific cytolytic functions. Proceedings of the National Academy of Sciences of the United States of BP Oil Synopsis Essay, America. 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 HIV infection. Role Of Iranian Woman Persepolis Satrapi Essay? J Clin Immunol. 2001; 21(3): 227-33.

Lambotte O, Ferrari G, Moog C, Yates NL, Liao HX, Parks RJ, et al. Heterogeneous neutralizing antibody and antibody-dependent cell cytotoxicity responses in HIV-1 elite controllers. AIDS. 2009; 23(8): 897-906. Dalgleish A, Sinclair A, Steel M, Beatson D, Ludlam C, Habeshaw J. Failure of ADCC to Synopsis, predict HIV-associated disease progression or outcome in 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. Essay? Antigenic specificity of antibody-dependent cell-mediated cytotoxicity directed against human immunodeficiency virus in antibody-positive sera. Synopsis Essay? J Virol. What´s A Friendship?? 1989; 63(2): 584-90. Doria-Rose NA, Connors M. Antibody-secreting B cells in BP Oil Spill, HIV infection. Essay? 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. BP Oil Synopsis? 2007; 5(6): 625-41. Sattentau Q. A Dissertation? Avoiding the 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 kill HIV-1- infected cells. J Immunol. Spill Synopsis Essay? 1994; 153(9): 4268-80.

Jolly C, Kashefi K, Hollinshead M, Sattentau QJ. What Is A Booty? HIV-1 cell to cell transfer across an Essay Env-induced, actin-dependent synapse. Role Of Iranian Satrapi Essay? J Exp Med. 2004; 199(2): 283-93. Groot F, Welsch S, Sattentau QJ. BP Oil Essay? Efficient HIV-1 transmission from macrophages to T cells across transient virological synapses. Blood. 2008; 111(9): 4660-3. Martin N, Welsch S, Jolly C, Briggs JA, Vaux D, Sattentau QJ. Virological synapsemediated spread of 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 multigenerational, 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 dont experience until their first post-doctoral experience. While at BP Oil Essay, NIH I will have received extensive support of staff from Richard Siegel, Director of the NIH MD/PhD partnership program and Michael Lenardo, Director the NIH/Oxford/Cambridge Scholars Program. What Is A Booty? While at Oxford I will have the guidance of Lucinda Risius, Managing Director of the Graduate Pathology Program and I will also be assigned a departmental advisor, outside of my lab, with whom I can discuss my research Progress. Educational Resources : Through the FAES at NIH I have access to courses to BP Oil Essay, improve lab specific skills and foundation knowledge.

I have already participated a Vaccine Development course. A Discussion Essay? At Oxford I will take seminars at the Centre for Excellence in Teaching and Essay, Learning for training in teaching skills. 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 what is a booty, scholarly research. Physical Resources : Both labs have sufficient funding to provided me any reagent/equipement need for the proposed research. Lab Space I am provided with a desk and personal desktop computer. Bench space is shared within the lab. BP Oil Synopsis Essay? Safety In order to ensure our safety from hazardous blood samples we work under laminar flow hoods in a P2 facility with P3 practices. 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 high international regard for its strong teaching and writing, research background and its unique and BP Oil Spill Synopsis, personal teaching style.

Within the What´s a Friendship? Essay, University the 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 BP Oil Essay, which each student has a principal supervisor, the possibility of a cosupervisor, a departmental advisor, and a college advisor. They By Tim O’Brien? The supervisor(s) will generally meet with the student multiple times a term, and each of the non-supervisory mentors will meet with the candidate approximately once a term and will file reports on the candidates 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 internal seminars from leading and more junior University and Departmental investigators and students. Adjoa will attend and participate in the weekly lab journal club, in BP Oil Synopsis, 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 other reading and what is a booty, writing. Adjoa will have her own writing up space close to BP Oil Spill Synopsis, her work bench, equipped with a portable computer and if required, an external screen. Adjoa will have full access to Role Woman Persepolis Satrapi, 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 Spill Synopsis Essay, biochemistry and chemistry departments and the Weatherall Institute of Molecular Medicine at 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 ancillary equipment that is 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 The Things They Carried Essay, 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 Spill Synopsis Essay, resolution, two of Persepolis by Marjane, which are set up for Spill Synopsis Essay live cell immunofluorescence work. 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 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 a skilled dedicated operator. Our collaborator in writing a dissertation, Oxford (Kay Grunewald) is located at the University Churchill campus, which is 4 miles from the Pathology Department. He maintains and BP Oil Spill Synopsis Essay, 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 The Things They Carried O’Brien Essay, this sample application can't reproduce the layout of the Spill Synopsis Essay, actual PDF forms. See below for the biographical sketch attachments.]

My interest in studying infectious diseases stems from the Essay, 2001 anthrax scare in Synopsis, the Washington D.C. area. Role Of Iranian Persepolis By Marjane Essay? I already had a strong interest in science but through my participation in the Gene Search Science Project at Catholic University the following summer I came to understand how medical research impacts treatment for routine illnesses and Essay, also plays a role in national defense. Since middle school I have been preparing for a career in science. Writing? 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 BP Oil Spill Essay, 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 multigenerational transmission, 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. My lab skills and BP Oil Spill Synopsis Essay, understanding of a research career increased exponentially during my four years in Michael Summers lab. Not only did I learn about protein purification, gel electrophoresis, NMR, and what is a booty, ITC while studying MuLV RNA packaging, but more importantly of the Spill Essay, time and discipline necessary for What´s a Friendship? 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 BP Oil, Tom Cech I had the responsibility of designing a new protocol to They Carried O’Brien Essay, study telomerase assembly, a skill that has enabled me to design a novel assay to investigate AIDS specific questions in my graduate studies. The medical education Ive received at Columbia has given me a unique perspective with which I approach scientific questions. Essay? Under the tutelage of Role of Iranian Woman in Book, Satrapi, Mark Connors and BP Oil Spill Synopsis Essay, Quentin Sattentau, leaders in their fields on nonprogressive HIV infection and viral transmission, my understanding of infectious disease and multigenerational transmission process, my capacity to pursue further research in these fields will be greatly enhanced. BP Oil Spill Synopsis Essay? In addition, the courses, medical preceptorships, presentations, teaching, and career development activities I am undertaking in the forthcoming years make me an what is a booty 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. BP Oil Spill Synopsis Essay? Composition and sequence-dependent binding of RNA to the nucleocapsid protein of Moloney murine leukemia virus. Biochemistry. 44(10), 3735-3744. Of Iranian Persepolis Satrapi? 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. 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. BP Oil Spill Essay? 2003. Biosynthesis of 2,3-diphosphoglycerate in Erythrocytes. Poster presentation at what is a booty, ERHS Science Fair and BP Oil Essay, Prince Georges County Science Fair. March 2003.

Smalls-Mantey A, Dey, A, Phillip C, Summers MF. Multigenerational Transmission? 2004. Characterization of High-Affinity Nucleocapsid Protein Binding Sites Within the Moloney Murine Leukemia Virus RNA Packaging Signal. Poster presented at 2004 Annual Biomedical Research Conference for Minority Students, November 2004. BP Oil Synopsis? Smalls-Mantey A, Min J, Pavletich N. 2005. Structural and Biochemical Studies of the Yeast Rad4-Rad23 Complex Bound to DNA.

Poster presented at Tri-Institutional Gateways to what is a booty, the Laboratory Program Symposium and 2005 Leadership Alliance. Spill? August 2005. Smalls-Mantey A, Miyazaki Y, Summers MF. 2007. Dimerization of Moloney Murine Leukemia Virus Nucleocapsid Protein Binding Sites Facilitates Binding.

Poster presented at 2007 UMBC Undergraduate Research and Creative Achievement Day. Writing A Dissertation? April 2007. 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. Spill? 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 What´s Essay, 2010 NIH/Oxford/Cambridge Scholars Research Colloquium. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. 2010. BP Oil Spill Synopsis Essay? HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells. Poster presented at Role of Iranian Persepolis by Marjane Satrapi, 2010 International AIDS Vaccine Conference. September 2010. Smalls-Mantey A. 2010.

Antibody-Dependent Cellular Cytotoxicity and the Control of Spill Essay, HIV. 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 What´s a Friendship? considered one of the very top priorities in the field of HIV-specific immunity. Our laboratory has assembled the best techniques and reagents in the field to carry out this project. We have extensive experience in HIV-specific antibodies and BP Oil Synopsis, cytotoxicity. In addition we have a long track record of training students to prepare them for a dissertation a carreer in biomedical research. 1985 - 1986 Intern in Pediatrics, The Boston Floating Hospital, New England Medical Center, Boston, Massachusetts 1986 - 1988 Resident in 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 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 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 Allergy and Infectious Diseases, National Institutes of Essay, Health, Bethesda, MD January 1994 - June 1994 Clinical Fellow in Infectious Diseases, The Childrens Hospital of Philadelphia, Philadelphia, PA. June 1994 - December 2004 U.S. They Carried By Tim O’Brien? Public Health Service, Medical Officer, National Institute of BP Oil Spill Synopsis Essay, Allergy and Infectious Diseases, Clinical and Molecular Retrovirology Section, Laboratory of Immunoregulation, National Institutes of a dissertation, Health, Bethesda, MD December 2007- Present Chief, HIV-Specific Immunity Section, Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD. Recent Honors and Editorial and BP Oil Spill Essay, Review Board Activities. 2009 - U.S.

Public Health Service Outstanding Service Medal 2009 - present Vaccine Production Development Team Scientific Advisory Board, University of Role of Iranian Woman in Book, by Marjane Satrapi, 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 Essay highly associated with restriction of What´s a Friendship? Essay, virus replication in a subgroup of HIV infected long term nonprogressors. 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 BP Oil Spill Synopsis coupled to perforin expression and 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 HLA B*5701+ long-term nonprogressors and progressors to writing, restrict HIV replication is not caused by loss of recognition of Spill Essay, 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 by Marjane exogenous IL-2. BP Oil? 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 paracrine IL-2 requirement, CCR7 expression, frequency, and cytokine secretion, of What´s Essay, HIV-specific CD4+ T cells are a consequence of antigen load. BP Oil Spill Synopsis? J Virol 2007; 81;2713-2725. They O’Brien? 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, OShea 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 Spill Synopsis HIV-Infected Cell Elimination Associated with Immune Control. The Things Carried Essay? 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. Defective human immunodeficiency virus-specific CD8+ T-cell polyfunctionality, proliferation, and cytotoxicity are not restored by antiretroviral therapy. BP Oil Synopsis Essay? 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. Writing? Breadth of BP Oil Synopsis, 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. 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 what is a booty, Mascola JR. Rational design of BP Oil Spill, envelope identifies broadly neutralizing human monoclonal antibodies to HIV-1.

Science 2010 329:856-861. A Friendship?? Migueles SA and Connors M. Long-term nonprogressive disease among untreated HIV-infected individuals: clinical implications of understanding immune control of HIV. JAMA 2010 304:194-201. [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 Spill Essay, HIV. What Is A Booty? 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 BP Oil Essay, the area of structure-function relationships of writing, HIV-specific antibodies. Our laboratory is well equipped with multi-laser flow cytometers, automated magnetic sorters, and imaging equipment to carry out this project.

Our laboratory is supported by Essay the research budget of the Division of Intramural Research. Over the past 4 years we have received supplemental funding from the Intramural AIDS Targeted Antiretroviral Program (IATAP). This will continue for the next 2 years. We have also received funding from the Office of AIDS Research for 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 Research), Centre dImmunologie, Marseille, France 1999 - 2003 Reader, Section of what is a booty, Infectious Disease, Imperial College London, London UK 2003 - Professor of immunology, Department of Pathology, University of Oxford, Oxford UK. Other Experience and Spill Essay, Professional Memberships. 1995 - Member, American Association for the Advancement of 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 Pasteur Institute Paris France Scientific Evaluation Committee. Writing? 2005 - Member of the Vaccine Research Center (VRC) NIH Board of Scientific Councilors 2007 - Member of BMGF Vaccine Monitoring Centre SAB 2008 - Member of Spill Synopsis, NIH HIVRAD/Novartis Neutralizing Antibody Vaccine Consortium SAB 2009 - Member of study section for Woman Persepolis by Marjane Essay Swedish Program for International Development 2010 - Member of NIH study section for the B cell Immunology Partnership Program for 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. Essay? (2010) Virological synapse-mediated spread of human immunodeficiency virus type-1 between T cells is sensitive to what is a booty, entry inhibition. J. BP Oil Spill Synopsis Essay? Virol. 84: 3516-3527. Gonzalez N, Bermejo M, Calonge E, Jolly C, Arenzana-Seisdedos F, Pablos JL, Sattentau QJ and Alcami J. Multigenerational Process? (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 Sattentau QJ. (2008). Efficient HIV-1 transmission from macrophages to 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. Nature Cell. Spill Synopsis? Biol. 10: 211-219. Jolly C and Sattentau QJ (2007).

HIV-1 assembly, budding and 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. A Friendship?? Jolly C, Mitar I and Sattentau QJ (2007). BP Oil Essay? Requirement for an intact actin and 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. Virol. A Friendship? Essay? 2005 79: 12088-12094.

Jolly C, Kashefi K, Hollinshead M and Sattentau QJ (2004) HIV-1 cell-to-cell transfer across an Env-induced, actin-dependent synapse. J. Exp Med. 199: 283-193. Additional recent publications of Spill Essay, important to The Things They Carried O’Brien Essay, 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 BP Oil Spill, an anti-inflammatory environment. PlosOne 6: e15861. 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 a discussion essay, HIV-1 envelope glycoprotein antigenicity and immunogenicity. J. Spill Synopsis Essay? Mol. Biol. Aug 25th epub. Krashias G, Simon K, Wegmann F, Kok WL, Ho LP, Stevens D, Skehel J, Heeney JL, Moghaddam AE and Role of Iranian Persepolis by Marjane Essay, Sattentau QJ. (2010).

Potent adaptive immune responses induced against HIV-1 gp140 and influenza virus HA by a polyanionic carbomer. Vaccine 28: 2482-2489. Zanetti G, Briggs JAG, Grunewald K, Sattentau QJ and Fuller SD. SIV spike glycoprotein structure in BP Oil Synopsis Essay, situ determined by writing cryo-electron tomography. Plos Pathogens August 2 (8): e83 Sheppard NC, Bates AC and Sattentau QJ (2007). A functional human IgM response to 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 BP Oil Essay, layout of the a dissertation, 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. Antibodies (Abs) induced by Synopsis Essay 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. Transmission Process? 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 BP Oil Spill Synopsis Essay, 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 the target. We want to study ADCC in infected patients to understand the Role Woman in Book, Persepolis by Marjane, magnitude and 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. 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 Spill Synopsis, Fc type.

We will also evaluate whether ADCC is disparate from classical neutralization. What´s A Friendship? Essay? Finally we will use microscopy to Spill Synopsis Essay, 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 The Things O’Brien a function that has been shown to mediate protection from lentiviral infection. We hypothesize that variations in ADCC activity of sera are dictated by BP Oil Spill Essay the amount, specificity, and Woman in Book, Persepolis Essay, subclass of HIV-specific antibodies. Aim 1 : Characterize the BP Oil Spill Essay, potency of sera of writing, HIV-infected individuals in ADCC. In ADCC, Abs bind viral epitopes that are presented by BP Oil Synopsis Essay infected CD4+ T cells. NKs expressing an Fc receptor bind the Fc domain of the Ab and use perforin to deliver granzymes to transmission process, the HIV-infected cell.

Subsequently, granzymes induce apoptosis within the cell. Our lab has developed a flow cytometric assay that measures granzyme B delivered to Synopsis Essay, an HIV-infected CD4+ target T cell. A Dissertation? We will classify ADCC by 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 serum of HIV+ individuals with various rates of progression and BP Oil Spill Synopsis Essay, viral loads to determine which contain Abs capable of mediating the highest levels of ADCC. Compare the what is a booty, ADCC and neutralizing activity of patient sera. Aim 2 : Characterize the specificity and Synopsis, breadth of The Things O’Brien, 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 specific epitopes is required for ADCC. Define the Synopsis, breadth of the polyclonal sera by its ability to mediate ADCC in what is a booty, CD4+ T cells infected by Synopsis Essay 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 what is a booty, cryo-electron microscopy (cryo-EM) and tomography, we will examine the synapse formed between NK and other cells with potential ADCC activity (macrophages and neutrophils) and Synopsis Essay, infected target cells. We will specifically investigate: The structure of Role in Book, Persepolis Essay, a functional ADCC synapse. The kinetics of BP Oil Synopsis Essay, ADCC function in real time and its relation to antibody type and a dissertation, specificity. A role for antibody-dependent cell-mediated phagocytosis (ADCP) in elimination of HIV-infected cells. d. BP Oil Synopsis? Receptors and effector molecules central to ADCC activity against HIV infected cells. Since being identified as the causative agent of AIDS in 1983, HIV infection has reached epidemic proportions with an estimated 2 million people worldwide dying from 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 a dissertation imperative need for a vaccine.

One of the Synopsis Essay, most critical areas of research on vaccines for HIV is the search for a discussion correlates of immunity. At present the Spill Synopsis Essay, only method for determining the potential benefits of a vaccine with regard to protection from infection or lowering of a discussion, viral load upon infection remains very large clinical efficacy trials. Synopsis Essay? Determining correlates of a Friendship?, 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 BP Oil Spill Synopsis Essay, immunity to transmission, HIV have largely focused on BP Oil Spill Essay 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 a discussion essay infected target cells and 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 Synopsis Essay, targets 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 of Iranian Woman Persepolis Satrapi Essay, colleagues demonstrated that neutralizing Abs with mutated Fc? receptor binding regions were less effective in protecting macaques from Spill Essay, 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 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. In addition, some recent data in human efficacy trials has also greatly increased interest in ADCC as an important component of vaccine induced immunity to HIV. In September 2009 data from the phase III RV144 (Thai) vaccine trial were released and showed that the prime-boost combination of What´s, ALVACR HIV and AIDSVAXR B/E reduced HIV infection. BP Oil Spill Synopsis? This vaccine regimen was known to multigenerational transmission process, 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 BP Oil Synopsis likely mediated through ADCC 5. Our laboratory is a discussion part of a large international effort to characterize ADCC antibodies from this trial in order to determine whether there were higher ADCC-mediating antibodies in Spill Synopsis, uninfected vaccinees compared to They O’Brien, 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 number, affinity, and specificities of Spill Synopsis, antibodies necessary to mediate ADCC remains poorly understood. Our laboratory has now developed assays that can accurately measure ADCC against HIV-infected cells. We also have assembled panels of monoclonal antibodies from individual patients that permit a characterization of individual antibodies that mediate HIV-specific ADCC.

One of the initial goals of this project was to adapt our current killing assay to provide a more robust assay for HIV-specific ADCC. They O’Brien? Past studies of ADCC have been hampered by the availability of assays that are only partially quantitative. There are four primary methods that people use to Synopsis Essay, measure ADCC 6. The most prevalent is the chromium release assays (CRA), which is transmission highly variable, does not specifically account for the mechanism of killing, and BP Oil Synopsis Essay, is insufficiently quantitative 7. The ADCVI assay incubates targets, serum, and effectors for 24 hours and a dissertation, the supernatant is applied to the B95-8 cell line for 5-7 days. The endpoint is reduction of viral infection of the B95-8 cells 8. This method measures inhibition of Synopsis, viral production and subsequent infection but its disadvantages lie in the length of the assay and failure to measure killing of infected cells. Another assay is the What´s a Friendship?, intracellular cytokine staining ADCC (ICS-ADCC) assay measures IFN-?, the Essay, primary cytokine responsible for the antiviral activity of many cells including NKs, CTLs, Th1, and dendritic cells 9. The Things They By Tim O’Brien? The advantages of this assay are that multiple effector functions can be measured and 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 a sensitive method, it likely places artificially high amounts of protein on the surface of infected cells, potentially overestimating the efficacy of BP Oil Synopsis Essay, 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. Role In Book, Persepolis Satrapi Essay? This is an adaptation of an 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 produce accurate quantitative and Synopsis, highly reproducible results. What Is A Booty? In brief, CD4+ T cells are stimulated for 3 days and HIV infected. Autologous, negatively selected NKs are used as effectors. 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 Synopsis, the target cell and What´s a Friendship?, 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). Synopsis Essay? Some infected cells show downregulation of CD4 (PE-Cy7). A reduction in the number of p24 stained cells is Carried by Tim O’Brien Essay represented as Infected Cell Elimination (ICE) and is an indication of Spill, cell death. In the example above, ICEInfected+Effectors+Serum = [(58.2-25.8)/58.2] x 100 = 54.9%.

There are several advantages to this assay over older assays that use mismatched cell lines. Because NKs lyse target cells in the context of MHC mismatch and cause nonspecific killing, autologous cells are used in our assay to Woman in Book, Satrapi, reduce background. 12. In addition, because NK function is diminished in HIV-infected patients, we use the NK cells and Essay, CD4+ T cell targets from writing, uninfected patients in our assay as a readout for the effectiveness of Essay, a given serum sample in mediating ADCC 13, 14. We observed variations in NKs from normal individuals in essay, their ability to mediate ADCC and choose donors that provide optimal signal to noise ratio that we will use to compare the serum of Synopsis, HIV+ individuals (Figure 2). The targets used in a discussion, this assay are likely closer to the in vivo situation given autologous HIV-infected targets are used in the place of antigen pulsed cell lines.

Other advantages include our assays ability to measure the mechanism of ADCC (granzyme delivery) and the outcome (death of infected cells) (Figure 2). MHC-1 is down-regulated in BP Oil Synopsis Essay, virally infected cells, and the loss of MHC-mediated killer inhibitory signals permit NKs to kill a cell 15, 16. A Dissertation? We control for Synopsis this by O’Brien measuring the granzyme content of Synopsis Essay, targets that are incubated only with effectors and a Friendship?, 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 BP Oil Spill, magnitude of ADCC in HIV infected individuals . Rational : There is considerable controversy regarding the magnitude of ADCC activity in What´s Essay, 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 BP Oil, disease progression 19, 20.We will compare the serum of HIV+ individuals with various rates of progression and viral loads to determine which contain Abs capable of mediating the highest levels of ADCC. We will also compare the The Things Carried by Tim Essay, ADCC and neutralizing activity of patient serum to evaluate whether or not ADCC and neutralization always occur together in polyclonal sera which is yet to be determined 21, 22. Methods : Over the past several years, in BP Oil, collaboration with the Vaccine Research Center (VRC) our laboratory has focused upon 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 immunity with ADCC. Expected Outcomes/Interpretation : Based on in vitro and of Iranian Woman Persepolis Essay, 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 BP Oil Synopsis, specificity. Rational/Methods : HIV-infected cells express Envelope (Env) protein on their surface which is recognized by NAbs and what is a booty, presumably by ADCC-mediating Abs.

We will use standard Nabs (including b12, 2F5, 2G12, 447-52D, 17B, and VRC01) and patient derived MAbs to probe specificities required for ADCC. An ideal HIV vaccine would be able to protect against 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 Synopsis different clades. We will use multiple strains of virus from clade A/E, B, and C to infect CD+ T cells and compare killing of target cells in the presence of serum pooled from clade B HIV+ patients. IgG1 and IgG3 are the a Friendship?, 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 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 Synopsis the surface of an infected cell in The Things, their natural conformation. Expected Outcomes/Interpretation : Determining the association between ADCC activity and Spill Essay, IgG subclass may provide insights into the necessity of What´s Essay, eliciting a specific class of IgG via particular antigens and adjuvants. Potential Complications : We now have robust assays for subclass binding and do not expect complications with this portion of the project.

Aim 3: Evaluation of the target-effector synapse. Rational : Using microscopy to BP Oil Synopsis Essay, 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 anti CD3 heavy chains that would redirect CD8+ T cells to a discussion essay, 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 targets were conjugated and undergoing lysis and by Spill 24 hours all targets had been lysed. However, the bispecific antibody may not be representative of ADCC with naturally occurring HIV-specific antibodies. In addition, the structure of the Role of Iranian Persepolis, 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 nanotube structures for viral transmission between CD4+ T cells 25. Spill? 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 ADCC in real time we will use time-lapse laser scanning confocal microscopy (time-lapse LSCM). Cell populations will be negatively selected from 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 The Things They Carried, evaluate ADCC; or neutrophils/monocytes to evaluate antibody-dependent cell-mediated phagocytosis (ADCP) and applied to poly-L-lysine coated cover slips. Cover slips will be fixed in paraformaldehyde, quenched with NH4Cl, permeabilized with Triton-X, and BP Oil Essay, stained for granzyme B, IgG, CD4/CD16/CD14 with fluorescent probes. Transmission? 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 BP Oil Synopsis, tomography (TEM) to generate higher resolution static images of the ADCC functional synapse. For TEM targets, serum, and effectors will be incubated for approximately 1 hour, fixed in paraformaldehyde and Woman in Book, by Marjane Satrapi Essay, glutaraldehyde, and stained with osmium tetroxide. BP Oil Synopsis? Samples will be embedded in epoxy resin, cut, and essay, imaged using a 300kV FEI Technai TF30 27. BP Oil Spill Synopsis? If necessary cryo-EM will also be used for what is a booty structural studies with the aid of Oxford collaborator Kay Grunewald. These experiments will be completed using the pathology departments core bioimaging facility and I will take departments Advanced Light Microscopy and Electron Microscopy course in preparation for BP Oil this research. Expected Outcomes : We expect to elucidate the The Things O’Brien, 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. BP Oil? 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 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 our large collection of cells, sera, and detailed clinical data from patients that are part of the NIAID HIV clinic, and both labs extensive experience in this area.

Samples are collected under the NIAID IRB-approved protocol 02-I-0086. Protections for human subjects are in accordance with NIAID Division of transmission, 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 BP Oil Synopsis Essay, 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 writing 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 BP Oil 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 staff will provide. In addition, on very rare occasions (less than 1 in every 1,000 times), malfunction of the leukapheresis machine can result in writing, 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 Race Consideration. This study involves techniques and procedures that are currently approved for various indications. BP Oil Spill Essay? 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 the NIH Clinical Center. Patients will be recruited from outside the NIH community by advertisements posted in the Clinical Center, the NIAID web site, and what is a booty, advertised in other media. Essay? We will attempt to facilitate minority recruitment using existing NIAID community outreach efforts involving local clinics in 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 the opinion of the what is a booty, study team, will not be offered participation in the study. Patients stopping HAART therapy will be monitored closely for virologic and immunologic relapse and BP Oil Essay, will be offered therapy, based upon the algorithm for essay 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 BP Oil Essay, HIV-infected adults greater than or equal to 18 years of age.

As an a discussion essay observational study there is no therapeutic benefit or clinical intervention resulting from participation in BP Oil Synopsis Essay, 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 Role of Iranian Woman Persepolis by Marjane Satrapi Essay, make valid comparisons within that cohort. The potential risks of withdrawing therapy in minors have not been studied and therefore may present unknown additional risks to Synopsis, that study population. Comparable units specializing in 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 writing a dissertation, nursing, and current guidelines continue to advise HIV-positive women to take antiretroviral therapy to prevent viral transmission to BP Oil Essay, the child during that time period. Therefore, pregnant or nursing women will be excluded from this study. What Is A Booty? Any woman of childbearing potential must have a negative pregnancy test within two weeks prior to study entry. Essay? All participants who engage in heterosexual intercourse must use two forms of in Book, Persepolis, reliable birth control for the duration of the study. When I entered the NIH/Oxford/Cambridge Scholars program I identified Mark Connors and 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 they agreed to start a collaborative project with me. After studying NAbs with Mark Connors for one summer I decided I wanted to focus on BP Oil antibodies that mediate ADCC.

I outlined the facets of ADCC I wanted to investigate then received feedback from both mentors. I subsequently wrote the O’Brien Essay, proposal that was edited by 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 Synopsis Essay, plan future experiments. We also conduct quarterly phone conferences with Dr. Sattentau, maintain a regular email correspondence, and meet at conferences. Communication will continue in essay, the same manner while I am in Spill, Quentin Sattentaus lab.

Selection of Sponsors and Institution. Selection of Institution. I decided to join Columbias 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. Role Woman Persepolis Satrapi? I decided to join the NIH/Oxford/ Cambridge Scholars Program to Synopsis Essay, complete my dissertation research because it would provide me access to transmission, 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 Synopsis, the pathology department I am allowed to select seminars and accelerated 1-day to 1-week courses that allow me to attain skills quickly and a discussion, immediately implement them into my research. In addition, the NIH/Oxford/Cambridge Scholars program provides funding for students to BP Oil Spill Synopsis, take courses offered by what is a booty any institution giving me access to virtually any educational opportunity necessary. In addition, through NIHs Foundation for Advanced Education in the Sciences Ive had the opportunity to take courses that will help me towards my goal as in infectious disease investigator such as the BP Oil Synopsis Essay, Vaccine Development course I completed.

The NIH/Oxford/Cambridge Scholars Program is designed so that students work with a mentor at a discussion, NIH and one at a university. When I joined the program I searched for HIV investigators throughout NIH and BP Oil Spill Synopsis, Oxford and identified Mark Connors and Quentin Sattentau as potential mentors because I wanted to conduct research that had direct implications in vaccine development. A Dissertation? Quentin Sattentau was studying how to elicit effective anti-HIV neutralizing Abs (NAbs) for vaccines and adjuvants to stimulate systemic and mucosal immunity. Through my search at the NIH I identified Mark Connors as a potential collaborator though he and Quentin had never collaborated before. I believed Dr. Connorss expertise in NAbs of Synopsis Essay, long-term nonprogressors (LTNPs) would complement Dr. What Is A Booty? Sattentaus vaccine development goals.

In addition, Dr. Connorss unique cohort of LTNPs would provide ready access to BP Oil Spill Synopsis, samples to study many facets of The Things Carried by Tim Essay, HIV infection. Finally, I believe Dr. Sattentaus approach to Synopsis, research as a PhD and Dr. Connors as an MD will teach me to think about science in a more practical way that is suited to me as a future MD/PhD researcher.

I will follow the principles on Woman Persepolis Satrapi responsible conduct of 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 BP Oil Essay, understand the responsibility scientists have to one another and medical community honestly share information. As a Columbia medical student, I have taken the Hippocratic Oath to guide my patient care and what is a booty, I am expected to adhere to the code of BP Oil Essay, ethics as outlined in student handbook. Upon joining the What´s a Friendship?, 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 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 a patient confidentiality seminar. Oxford offers an Ethics: Confidentiality course in which I will enroll to better my further my understanding and ability to maintain ethical practices with regards to research.

As a physician, I hope to alleviate an Synopsis individuals immediate suffering; and as a scientist I hope to contribute to long-term solutions that can prevent suffering. To work most effectively at writing a dissertation, the interface of medicine and science, I have endeavored to earn an MD/PhD. I believe that the BP Oil Spill Synopsis, combined degrees greatest value lies in the perspective that it affords: to approach patient care with a researchers eye, and to approach research with a physicians focus on improving the of Iranian Woman Persepolis Satrapi Essay, quality of life. A few years ago I conducted research on Synopsis Essay telomerase, an enzyme that maintains healthy chromosome length. Role Woman By Marjane Satrapi? When telomerase is upregulated, as it is in 90% of 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. Synopsis? Secondly, these cells had abnormal morphologies, as do tumor cells. A Discussion? 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 any substantial research concerning this question. Research in biochemistry affords many new advances in medicine, but through that experience the Spill Synopsis, importance of keeping a broader medical picture in a Friendship? Essay, mind became clear to me. My goals as a physician scientist are to treat patients and BP Oil Spill Essay, 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 what is a booty, infectious disease. The training I will receive in Oxfords doctorate program will prepare me to BP Oil Spill Synopsis Essay, 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 even through this grant. These are all specific skills that would be harder and The Things They Carried by Tim, take longer for me to garner through a traditional medical student experience. The advantage I have in understanding that I want to 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 Connorss 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 Essay, week with Dr.

Connors. We also have phone conferences and meet with Dr. Sattentau at numerous conferences throughout the year. Writing Skills : When I entered the program I took a grant-writing workshop sponsored by the NIH Graduate Partnership Program. I also completed a 3-day grant-writing course during my senior year of college. I will improve my writing skills through preparation of a discussion essay, manuscripts based on my research to be submitted for publication, applying for Spill Synopsis Essay training grants, and submission of Essay, a report for my transfer viva (qualifying exam). I will also complete the BP Oil Synopsis, Scientific Writing from a Readers 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 process the American Psychiatric Association so I could learn to Spill Synopsis, effectively interact with HIV+ patients I encounter. A Friendship? Essay? I attend weekly seminar series hosted at the NIH such as the Wednesday Afternoon Lecture Series, Immunology Interest Group seminar series, and BP Oil Essay, 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 pertinent Keystone Symposia and multigenerational transmission, 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 my graduate program journal club and will have more opportunities to present my research in lab meetings.

Career Development : The NIH/Oxford/Cambridge Scholars programs hosts a seminar series where leaders in government/academic research, the pharmaceutical industry, and Spill Essay, finance speak with us about their career. I also attended NIAID Fellows retreat that focused on a Friendship? career development. Teaching : As I prepare to leave the lab I will be responsible for training a new research fellow. Year 1 (continued) and BP Oil Essay, Year 2: Quentin Sattentau Lab (Oxford) Research 75%, Seminars/Courses 10%, Presentation/Career Development/Clinical 10%, Teaching 5% Research Skills : While working in Quentin Sattentaus 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 year. Essay? I have also been assigned a graduate advisor within the pathology department but not in BP Oil, 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 They Carried 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 Essay, Statistics. Bench to 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 transmission, be an undergraduate lab instructor and take the Oxford course Teaching and Synopsis Essay, Learning Skills to prepare myself. Year 3 and Year 4: Medical School (Columbia) In the summer after 11th grade I participated in what is a booty, the intensive 6-week Gene Search Program at the Catholic University of America that introduced me to molecular biology and the exciting race to sequence the BP Oil Spill Synopsis, 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 The Things Carried O’Brien Essay, design experiments. Because of Spill, my accomplishments that summer, I was invited to What´s a Friendship?, participate in BP Oil Spill Synopsis, an extension of the Gene Search Program during my 12th grade year. This program inspired me to pursue a career in biomedical research because I began to The Things by Tim, understand how the principles I was learning in school fostered scientific discovery. As a high school senior, I worked with Dr. William Winter at Synopsis, Howard University School of Role Woman in Book, Essay, Medicine to investigate pathways that regulate 2,3-diphosphoglycerate (2,3-DPG) levels in red blood cells (RBC). 2,3-DPG lowers the BP Oil Spill Essay, 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 what is a booty, patients with sickle cell anemia, deoxygenating hemoglobin ultimately causes red blood cells to sickle by promoting hemoglobin polymerization.

Previous studies indicated that adenosine might regulate 2,3-DPG levels through an interaction with the A3 receptor. Synopsis Essay? To test this idea, we indirectly measured 2,3-DPG levels by writing using spectrophotometry to measure a by-product of the 2,3-DPG pathway. An exciting therapeutic prospect of our research is Spill Synopsis that an what is a booty 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 schools Science and Technology Program, and presented it at BP Oil Spill Synopsis, my schools 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 what is a booty key to developing gene and BP Oil Spill Essay, 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 Psi-site. Previous work in the Summers lab suggested a riboswitch mechanism for 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. Persepolis By Marjane Satrapi? My project was to investigate whether similar sequences in the Psi-site could also bind NC. Spill Essay? 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 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 guanosine. The paper I co-authored (Dey, A., et. al., 2005) extends an established model for genome recognition, in which the NC domains of assembling Gag molecules interact with multiple X(i-3)-X(i-2)-X(i-1)-G(i) elements (X is a discussion a variable nucleotide) that appear to be preferentially exposed in the dimeric RNA. BP Oil Spill Synopsis? 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 what is a booty, gel-shift assays. As expected from 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. Nikola Pavletich toward solving the crystal structure of the Saccharomyces cerevisiae Rad4-Rad23 complex bound to a DNA substrate. Rad4-Rad23 is BP Oil Spill a homolog of human XPC-HR23B, a DNA damage-repair protein. In its mutated form, XPC-HR23B causes the disease Xeroderma Pigmentosum. Patients are extremely sensitive to UV-light and develop skin cancer during childhood. I identified DNA substrates to Role of Iranian Woman Satrapi Essay, be used in crystallization trials by HPLC-purifying DNA, performing gel-shift assays, and setting up crystal trays. I presented this work in a talk before the Tri-Institutional community and Synopsis Essay, during posters sessions at Woman in Book, Persepolis Satrapi, the 2005 Leadership Alliance National Conference and at the 2005 SACNAS National Conference. In 2006 I studied telomerase assembly in the lab of Dr.

Thomas Cech at BP Oil, the University of Colorado at Boulder. The central role telomerase plays in protecting chromosome integrity places telomerase at Woman in Book, Satrapi Essay, the heart of BP Oil Essay, two common human ailmentscancer and aging. To study how and where in the cell telomerase is assembled from its RNA and essay, protein components, we used the Synopsis Essay, 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 that a polyadenylated precursor form of 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 the nucleus, where the a discussion essay, telomerase holoenzyme is assembled. Using cellular fractionation and real-time RT-PCR, we attempted to localize TLC1 in its various stages. During the summer between my first and 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 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 many nuances of 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 Synopsis, vital role that a physician-scientist plays in translating basic research into essay, medical treatment. BP Oil Spill Synopsis? Although Ive experienced frustrations and what is a booty, disappointment in research, the thrill of Synopsis, observing something new, finishing a project, or publishing a paper always outweighs these challenges. What Is A Booty? Each successive experience only Spill Synopsis Essay increased my eagerness to improve health care through innovative research. [Redacted from sample.] QUENTIN SATTENTAU MENTOR STATEMENT. [Redacted from sample.] [Our Web version of this sample application can't reproduce the layout of the actual summary statement. See below for the data fields and a discussion, 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 extent of protection that ADCC provides. The reviewers noted that the candidate is outstanding, with a strong record of Synopsis Essay, research training and accomplishment. Transmission Process? 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. BP Oil Spill Synopsis? Minor weaknesses of the application included the ambitious research plan to be accomplished in two years, the The Things O’Brien Essay, risk of conducting the research at two sites, and the failure to address pitfalls and alternatives of the research project. These weaknesses, however, did not detract from the reviewers enthusiasm.

This training project will have a high impact in Synopsis Essay, ensuring that the candidate maintains her potential to What´s a Friendship?, become an independent clinical researcher. DESCRIPTION (provided by applicant): Understanding the basis of an immune response that controls infection or provides sterilizing immunity remains a major goal in the search for Spill Essay 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. What Is A Booty? However, this approach has not been successful in containing viral replication in Synopsis Essay, vaccinees that have become HIV-infected. Essay? Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against Spill, challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. What´s A Friendship? Essay? 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 BP Oil, the target. What Is A Booty? We want to study ADCC in infected patients to understand the magnitude and Spill Synopsis Essay, 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. Based on these findings, we will select the a discussion essay, sera of patients with the most ADCC, generate monoclonal Abs (mAbs), and Synopsis Essay, characterize the a discussion, mAbs based on epitope specificity, affinity, potency, breadth, IgG isotype, and BP Oil Spill Synopsis, Fc type. We will also evaluate whether ADCC is disparate from what is a booty, 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 BP Oil Synopsis, 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 a function that has been shown to mediate protection from The Things O’Brien, lentiviral infection. We hypothesize that variations in ADCC activity of BP Oil Spill Synopsis Essay, sera are dictated by the amount, specificity, and They by Tim O’Brien Essay, subclass of BP Oil, HIV-specific antibodies. Aim 1: Characterize the potency of sera of HIV-infected individuals in They Carried by Tim O’Brien Essay, ADCC.

Aim 2: Characterize the BP Oil, specificity and breadth of antibodies with ADCC activity. Aim 3: Characterize the essay, structure and function of the target-effector synapse using both fixed and live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and BP Oil Synopsis, cryo-electron microscopy (cryo-EM) and tomography. Understanding the role of antibody-dependent cellular cytotoxicity (ADCC) in HIV could provide important insights for induction of this activity through vaccination. This project seeks to characterize the What´s a Friendship?, Abs that mediate ADCC and image the functional synapse formed by cellular components involved in ADCC with the BP Oil Spill, goal of a dissertation, defining new goals for the development of HIV vaccines and therapeutics. Fellowship Applicant: 1. Sponsors, Collaborators, and Consultants: 1. Research Training Plan: 2. Training Potential: 1. Institutional Environment Commitment to Spill Synopsis, Training: 2. Overall Impact/Merit : This proposal is from an multigenerational 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 an NIH laboratory and in a university research lab. Ultimately, she will receive her Ph.D. from Synopsis Essay, Oxford University and complete her medical training at Columbia University. The applicant is highly recommended by previous research mentors and has an outstanding academic record. Her research experience, which includes four years of undergraduate research, is matched by Role of Iranian in Book, Essay her productivity; she is a contributing author on two published papers and has presented her work at numerous scientific venues. The research plan, which derives directly from the 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 Spill Synopsis, antibody subtype. 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 Things They Carried By Tim O’Brien Essay? The training plan is unique in Synopsis Essay, that training and research will be accomplished in two separate laboratories of the sponsor and co-sponsor. This proposal outlines an multigenerational process excellent training opportunity that will greatly benefit the applicant and her potential for establishing an independent research career. The applicant, who is currently enrolled in an M.D./Ph.D. 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 BP Oil Spill Synopsis, more than 200 current and writing, former undergraduates that he has mentored. The applicant has four years of Spill Synopsis Essay, undergraduate research experience, which involved work with the multigenerational transmission, Moloney Murine Leukemia Virus, and BP Oil, research performed during a summer program at the Howard Hughes Medical Institute focused on studies of telomerase. Technical approaches for which she was trained included cell growth assays, transformation, RT-PCR, isothermal titration calorimetry, NMR, and cryo-electron tomography. Through her prior research experience, the applicant has been included as a contributing author on two published papers and multigenerational, has presented eight posters and one oral presentation at various local, national, and international meetings. The applicant has an outstanding academic record (3.97 GPA) and Spill Synopsis, numerous awards and recognitions reflecting superior academic performance. The collaborative project outlined in the proposal was an outgrowth of a dialog initiated by the applicant between the sponsor and O’Brien Essay, co-sponsor. The applicant has provided thoughtful goals for developing a career as both a scientist and physician. 2. Sponsors, Collaborators, and Consultants: The applicants mentor and sponsor, who is currently Chief of the HIV-Specific Immunity Section in the Laboratory of Immunoregulation, NIAID/NIH, is an established and Essay, productive senior investigator with research support provided by writing a dissertation the NIH. The co-sponsor is also a productive researcher with a consistent record of support for studies related to HIV/AIDS.

The research interests and expertise of the sponsor and co-sponsor will provide excellent support for the applicants research and BP Oil Essay, training. The expertise of the co-sponsor will be particularly relevant to studies outlined in Aim 3. Carried By Tim O’Brien? The sponsor has trained an average of four students per Spill Synopsis Essay year over 20 years, suggesting an excellent training environment for the applicant. Similarly, the co-sponsor has trained nine Ph.D. They Carried? trainees since 2003. Synopsis? During her predoctoral fellowship, the a dissertation, applicant will be interacting with four post-baccalaureate students and BP Oil Spill Synopsis, one M.D. in what is a booty, the sponsors lab, and four Ph.D. trainees in BP Oil Spill Synopsis Essay, the co-sponsors laboratory. Writing? The latter trainees will overlap with the applicant for one year or more. These interactions will enrich the applicants training. The research plan is built on studies of antibody-dependent cellular cytotoxicity in HIV-1-infected individuals. BP Oil Essay? Experiments will examine the Role of Iranian Woman Persepolis by Marjane Satrapi, potency of sera of HIV-1-infected individuals in ADCC (Aim 1), the BP Oil Spill, specificity and breadth of antibodies with ADCC activity (Aim 2), and the structure/function of the target-effector synapse that mediates ADCC. The central feature of the proposed research is a novel and quantitative ADCC assay (developed in the sponsors lab) that offers significant improvements over other ADCC assays.

Results generated from experiments in Aims 1 and essay, 2 will have both basic science and translational value, while results from Aim 3 experiments will provide basic information about the mechanisms of Spill Synopsis, ADCC. The applicant and sponsors are well positioned to perform these studies, since the a discussion, patient populations and Essay, study procedures are already in place. The applicant is participating in the NIH/Oxford/Cambridge Scholars Program, which promotes research with a mentor at Role in Book, Persepolis by Marjane Satrapi Essay, 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 Spill Synopsis Essay, the sponsors lab at the NIH, continued studies and training in year 2 in the co-sponsors lab at Oxford, and years 3 and 4 in medical school. Training in transmission process, 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 Spill Essay presentation of results and 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 Role of Iranian Woman Persepolis by Marjane Satrapi Essay, scope of studies in BP Oil Synopsis Essay, Aim 1. Correlates of ADCC to be studied in Aim 1 (rates of Essay, progression and viral loads) were not described in sufficient detail. Synopsis Essay? 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 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 Role of Iranian in Book,, enhance the capability of the BP Oil Essay, applicant to Carried by Tim, develop an independent and productive research career. 5. Institutional Environment Commitment to Training: The resources and Essay, facilities at both institutions will provide outstanding environments for training and research. A more detailed plan regarding the coordination of training plans between the sponsor and co-sponsor would be valuable, considering the distance and a Friendship? Essay, time difference between the BP Oil Synopsis, two performance sites.

Training in 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 multigenerational, Oxford (both focused on ethics) is mentioned. Comments on BP Oil Spill Subject Matter (Required): Training courses and writing, 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 description of RCR training. Comments on Duration (Required):

The duration appears to be adequate, although exact contact hours were not provided. Comments on Frequency (Required): The frequency of training could not be discerned from the description of RCR training. Fellowship Applicant: 2. Sponsors, Collaborators, and 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 BP Oil 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. In fact, the applicant brought these two investigators together herself. Essay? Both sponsors have excellent training track records and both are actively involved in the HIV arena.

The primary goal of Spill Synopsis, this project is to characterize antibodies that mediate antibody-dependant cellular cytotoxicity (ADCC). What´s Essay? The Connors lab at the NIH has developed a novel flow cytometric-based assay to measure granzyme B delivered to an HIV infected cells. Using this novel assay the applicant plans to compare ADCC in HIV+ subjects with various rates of disease progression and to define the specificity and breadth of antibodies with ADCC activity (Aim 1 and Synopsis Essay, 2). The third Aim to characterize the essay, 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 Spill Essay, samples between NK and other cells with ADCC activity. If the applicant is successful the data from this project will be highly important for Role in Book, Persepolis HIV vaccine development. The research project however is Spill Synopsis quite ambitious and no timeline is given. It appears that the applicant plan to writing, spend 1 more year in the Dr. Connors lab and 1 year in BP Oil Essay, Dr. Sattentau lab making it a very busy two years if the applicant plans to transmission, finish all the work they outlined. Despite the highly ambitious nature of the BP Oil Essay, proposed research project the overall project goals warrant further study and the applicant herself is of the 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 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 high quality research and publications and a solid list of successful trainees (4/year for 20 years). The other sponsor, Dr. Sattentau, a Professor of Immunology and a discussion essay, Pathology at the University of Oxford, is equally meritorious with over 151 published manuscripts and solid funding record. BP Oil Spill Synopsis? Dr. Role Persepolis By Marjane Essay? 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 support the feasibility of the project. Spill Synopsis? The plan is to utilize a novel recently developed (Connors lab) quantitative flow cytometric assay to What´s a Friendship?, measure GrB cytotoxicity and Essay, determine which Abs are capable of writing a dissertation, mediating the highest level of ADCC. Using this technique the applicant plans to characterize the potency of sera from subjects with different rates of disease progression, characterize the specificity and breadth of Spill Synopsis, antibodies with ADCC activity. Role Of Iranian In Book, Essay? Using advance microcopy techniques (fixed and live cell laser scanning confocal microcopy and electron microscopy) the applicant plans to examine the structure and function of the target-effector synapse. This work will be done under the guidance of Dr. Sattentau who is an expert in studying the Spill Essay, virologic synapses. This is a highly ambitious grant. It may be difficult for essay the applicant to finish all the proposed work especially the Synopsis Essay, work proposed in Aim 3. The applicant is calling for 300 subjects but no power calculation is shown to 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. The plan includes research, writing skill (grant writing course), presentation, career development, teaching (both in class room training of new personnel) and discipline specific learning (i.e. in house courses and seminar series, lab meetings). What´s A Friendship? Essay? Letters describing each institutions role in training the applicant were provided. Applicant states that she will interact on Synopsis daily basis with section head and other senior laboratory member on The Things They Carried by Tim 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 Synopsis Essay, applicant will be at the NIH for one year and Oxford for one year and then back to medical school for the last 2 years. This raises some concerns regarding the multigenerational process, 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. Both institutions are of the highest caliber with access to BP Oil Synopsis, 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 Carried Essay, the Sciences also has specialized courses that will facility the applicants goal to become an Synopsis Essay infectious disease investigator. Training in the Responsible Conduct of Research: Comments on Format (Required): On line ethics course at a Friendship? Essay, the NIH and formal classroom course at Oxford. Comments on BP Oil Synopsis Essay 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 multigenerational transmission process 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 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 Spill part of the NIH/Oxford/Cambridge Scholars program, carrying out a discussion essay, her PhD research under the guidance of two excellent researchers and mentors, Drs.

Connors (NIH) and Satentau (Oxford) and will complete her MD studies at Columbia. The primary hypothesis of this proposal is that ADCC activity levels in sera are dictated by the amount, specificity, and Spill Synopsis, subclass of HIV-specific Abs. This represents a highly relevant area of research that will be tackled through 3 straightforward and 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 a discussion essay, 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 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 BP Oil Essay, relevant research in an excellent training environment.


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The Ultimate List of AP English Literature Tips. The AP English Literature and Composition exam is designed to test your ability to think critically and analyze literary excerpts. The test is three hours long and consists of a multiple-choice portion (worth 45% of your grade) and an essay portion (worth 55% of your grade). Here are some tips to help you get on BP Oil Essay, your way to making a 5 on the AP Literature exam. Before you start studying for what is a booty, the AP Literature exam at the end of the year, you need some tips on how to survive the course itself. Advance Placement (AP) courses are deliberately designed to BP Oil Spill Synopsis Essay, be more difficult than the standard high school classes; they are meant to challenge you. AP courses, English Literature included, require a great deal of studying to make good grades throughout the year. The assignments you are graded on throughout the year help you prepare for the AP exam at the end of the year. Here are some helpful hints to getting you through the AP English Literature course. 1. Transmission! Complete Any and Synopsis Essay, All Summer Work Assigned: AP Literature, as its title indicates, requires a lot of reading. Chances are, your teacher will provide you with a reading list and expect the required titles to be read when you walk into your first day of class.

In some cases, you may even be assigned a report or project to be completed before you begin the class. This is Role of Iranian Satrapi more for the teacher to view what literary skills you already possess and what skills will need to be taught to you. Spill Essay! However, this doesnt mean you should take the work assigned lightly. If you take it seriously and complete a proficient assignment, it will show your teacher that you are in the course to learn. This attitude will make the school year a lot more bearable for both you and your instructor.

2. Go to Class: Missing class leads to a discussion essay, missing material. Missing material leads to lower scores on assignments. Lower scores on assignments lead to lower scores on the exam at BP Oil Essay the end of the year. The bottom line is: dont miss class if you can help it. 3. Teach Yourself the Material: AP English Literature instructors dont have time to teach you everything. What Is A Booty! Since you are probably only in their presence for BP Oil Spill Synopsis, an hour or two, they have learned to prioritize the material they have to The Things They Carried, teach. Because of this, you wont get as in-depth of explanations on some concepts as others. Synopsis! You need to a discussion essay, learn how to Spill, teach yourself the material to really make the writing, most out of BP Oil Essay, this course. You can really get creative with this. You can teach yourself by conducting good old fashioned research, or just by reading the assigned texts.

Or, you can expand your knowledge a little more. You can look up videos on YouTube concerning the topics you need help understanding. You can also use to What´s a Friendship? Essay, test yourself on different areas covered in a typical AP English Literature class. 4. Learn How to Analyze Text: Analyzing literary text is an incredibly large portion of the AP English Literature course. Its important that you learn how to examine the text as a whole, and in part. Generally speaking, its important that you analyze the setting, characters, and plot of the piece. Spill! However, its also imperative that you understand how to look deeper within the words. Deconstruct the text and examine its theme, look for literary devices, and motives.

5. Read: This is literature! Therefore, you should be getting a good amount of reading done. This does not necessarily mean that you have to aim to read an outrageous number of books or anything. What Is A Booty! You just need to at least make an attempt to read every day. BP Oil Spill Synopsis! As you read, try to dissect the depth of the text. After a few days of this, youll be surprised at a discussion essay how easy analysis can come to you once you train your mind to question everything. 6. Ask Questions: Your teacher is there to BP Oil Spill Synopsis Essay, help; its their job. If theres anything you dont understand, be sure to ask your instructor. They Carried O’Brien! Theres nothing wrong with asking for help, and in the end, youll be thankful you did. Understanding a concept you previously had trouble with is Spill Synopsis sure to be a huge weight off of Role of Iranian Satrapi Essay, your shoulders.

7. Spill Synopsis! Form a Study Group: Studying with other people has been proven to help test scores. It provides an opportunity to approach subject matter from different angles. Some people in what is a booty, your group may know certain concepts better than you, and vice versa. One of the best ways to make sure you know the material is to teach it to others. 8. Synopsis Essay! Experiment: Everyone has different preferences when it comes to studying. Maybe youre a visual learner. Perhaps you like to listen to material to really understand it. The best way to find out what form of studying helps you best is to The Things They, experiment. Spill! Try different methods to see what works best for you.

Plus, keeping a variety in your study routine helps keep boredom at bay. Now that you have a grasp on how to get through the of Iranian Woman Persepolis by Marjane Satrapi, actual coursework of your AP English Literature and Composition class, its time to learn how to study for the exam at the end of the year. First, well take a look at some tips that are sure to help you ace the Spill Essay, first portion of the AP Literature exam: the multiple-choice section. This portion is worth 45% of your total score and it consists of several passages to read and 55 questions to answer, which you have exactly one hour to complete. Lets get started.

Start your AP English Literature Prep today. AP English Literature Multiple-Choice Tips. 1. What Is A Booty! First, Read the Passages: Yes, you read that correctly. Its a common misconception that you should read the questions before reading the passage, so you can work quickly and Essay, scan the a dissertation, text for the correct answer. One vital thing to remember is this: quicker isnt always better. On a timed exam, it is important to work at BP Oil Spill Essay a brisk pace, but do not move so quickly that you make simple mistakes. Its best to read the passages before even glancing at the questions because it prevents error. Writing! By merely scanning the passage for BP Oil Synopsis Essay, the answers, youre missing out on a plethora of content that could be vital to answering questions correctly later on in the test. 2. Look Deep Within the a Friendship?, Text: Its extremely important that you analyze the Synopsis, passages within the exam very carefully. Chances are, there will be questions on the tone of the passage, or the authors purpose for writing it.

Was it to inform or persuade the audience? Perhaps the essay, author used some literary devices like allusions or irony. BP Oil Spill Synopsis Essay! Closely read the passages and you will have no problem identifying the answers to a Friendship?, questions that are specific to the literature side of AP English. 3. Carefully Read the Questions: If you dont understand what the question is asking, you cant possibly expect to know the Spill Essay, answer. Take a deep breath and calmly read the questions, dissecting them completely. This will be easier to do for some questions than for multigenerational transmission, others. Once you understand what it is Synopsis Essay exactly the question is asking, try to recall where in writing a dissertation, the passage the answer could be located.

Also be sure to read the question in BP Oil Spill, its entirety. Sometimes, the writers of the test will throw in certain words or phrases that lead the question in a different direction. For example, the words EXCEPT and NOT are often used at the end of questions, and this can confuse you. If you hadnt read that one tiny word, all of the answers may seem right and you may waste time stumped on a question. 4. Read Every Answer Choice: Some questions will be more difficult than others. Some questions are even designed to trip you up. Be sure to Role of Iranian by Marjane Satrapi Essay, read every single word in every single answer choice; sometimes one word can make all the difference as to whether or not an BP Oil Synopsis Essay answer is correct.

5. Reread Parts of the Passage: If time permits, reread the parts of the text in which answers are located. Be sure the information matches one of the answer choices. You may even want to put a star, dash, or some other marking beside portions of the text that contain answers. That way, if you have extra time at a Friendship? the end of the Synopsis Essay, test, you can go back and check your answers more quickly. 6. Use Your Time Wisely: This is a timed exam. 60 minutes to complete 55 questions. This allows for an average of a minute per question, with some leftover time to a discussion essay, account for Spill Synopsis, reading passages. You have absolutely no time to sit at your desk staring blankly at questions you dont quite understand. Luckily, there is no penalty for answers marked wrongor answers not marked at a Friendship? Essay allon the BP Oil Synopsis Essay, AP English Literature exam. This means you should definitely skip the questions youre unsure of. Mark them in some sort of way so that it is noticeable that you havent answered them yet.

Then, if you have some time at the end of the transmission, test, you can go back and see if you can come up with the answer. Alternatively, if you cant seem to find an answer: guess! Remember, youre only graded on the number of questions you get right; theres absolutely no penalty for getting a question wrong. 7. BP Oil Spill Synopsis! Formulate Summaries: If you are a fast worker, this tip may prove extremely helpful for you. A few of the multiple-choice questions may test your overall comprehension of the passages you read.

In the margins of the page beside the passage, jot down a few bullet points outlining the plot progression. This way you can refer back to essay, your notes when answering questions rather than searching the entire text. 8. Synopsis! Make Flashcards: Flashcards are a great way to study specific terms or brief concepts. Since you will be tested on your understanding of certain terms, it is important that you know them like the back of your hand. Try making flashcards of different literary devices and review them periodically throughout the semester. 9. Study Everywhere: This may seem a bit extreme, but it really helps. Take the flashcards youve made with you wherever you go. Keep them in your wallet, in your purse, or even in your car.

Whenever you have a moment of free time, instead of scrolling through Twitter or Facebook on what is a booty, your phone, run through a review of Synopsis, your terms. Itll stick better in your memory and a discussion, help your AP Literature exam score in the long run. 10. Test Yourself: The most helpful and effective way to prepare for the multiple-choice portion of the AP English Literature exam is by testing yourself. Prepare early in the semester for the exam. BP Oil Essay! Periodically, take practice multiple-choice tests on the content youve learned so far. There are several websites out there dedicated to a discussion essay, helping you quiz yourself for the AP Literature exam.

One of these is , which allows you to test your abilities on nearly every concept covered in the AP English Literature course. 11. Dont Stress It: The AP English Literature exam is one big test. Sure, it affects the amount of college credit you receive coming out of high school. But at the end of the BP Oil Spill Synopsis Essay, day, its just a test. Anxiety and stress can severely affect your ability to function correctly. Over time, it can even start to have negative effects on your mind and The Things O’Brien Essay, body.

Some people even develop anxiety disorders. Just remember, your mental health is BP Oil Essay more important than your grades. Take a deep breath periodically throughout the test. Itll help calm your body and soothe your mind so you can concentrate better. Now that you have some tips on how to tackle the multiple-choice portion of the AP English Literature exam, its time to focus on the most challenging part: the free response portion. In this portion, you have two hours to complete three essays.

This section tests your ability to analyze passages and Role Woman in Book,, dissect them to form logical interpretations to be illustrated in your essays. Start your AP English Literature Prep today. Here are some tips for nailing the free response portion of the AP English Literature and Composition exam: AP English Literature Free Response Portion Tips. 1. Read the BP Oil Essay, Question: The first step towards writing an What´s a Friendship? Essay awesome essay on the AP Literature exam is BP Oil Spill reading (and understanding) the question. The Things They By Tim! What are the authors of the test asking for specifically?

The answer to this question is the key to writing a well-rounded essay. The scorers of the free response portion want essays that are clear and straight to the point. Simply restating the prompt will result in a huge deduction of points. Regurgitating the question will show the BP Oil Spill Essay, reader that you may not be confident in what is a booty, your ability to dissect passages. This is an extremely bad impression to give, since the interpretation of text is the basis of the entire course.

2. Come Up with a Thesis: A well-written thesis is the BP Oil Spill Essay, basis of a discussion essay, all successful essays. As mentioned previously, do NOT restate the BP Oil Spill Essay, question. Come up with your own unique topic sentence to answer the a dissertation, question. Spill Essay! Make it brief and to the point. You have the rest of the multigenerational, essay to elaborate on your thesis; that will be your body.

3. Stay Organized: Organization is Synopsis key to writing a great essay. Prepare an outline within the first few minutes of writing your essay. Perhaps even use a diagram, if youre a visual learner. A clear and precise outline can help prevent rambling when answering the question in your essay. 4. Use a High-Level Vocabulary: Since this is an exam for an Advanced Placement English course, it is imperative that you use a vocabulary that reflects a higher level of education. However, be sure that you use your impressive vocabulary in context. Nothing looks worse than using a word incorrectly in your essay. Be careful: only use words in in Book, Essay, which you know the BP Oil Essay, definition. 5. Use Your Resources: On the What´s, first two essays, you will be asked to read a passage and analyze it according to the instructions given in the question.

Use the passage to your advantage. Frequently refer back to specific parts of the text. This will show the BP Oil Synopsis, readers that you paid very close attention to detail when reading the passage. The specific references display the ability to close read, which is transmission process a skill covered frequently in an AP English Literature course. 6. Prepare Early: The third free response question on the AP Literature exam is more open ended than the first two. You will be asked a question and you will be given the opportunity to BP Oil Essay, answer it pertaining to a work of literature that you have read in class. Its important that you keep this particular essay question in mind as you work throughout the semester.

If a particular work of literature stands out to you, prepare early to multigenerational transmission process, choose this as the piece to write about in your third essay. 7. Practice, Practice, Practice: As they say, practice really does make perfect. BP Oil Essay! A good option for practicing free response questions involves searching the Internet for old exam rubrics. These show you exactly what the scorers are looking for of Iranian in Book, by Marjane, in an essay. The AP Literature section of AP Central, a website created by the College Board to help with studying for exams, has several practice exams for Spill Synopsis, your use. Take advantage of this and practice writing essays using different prompts from previous exams. 8. Use a Good Writing Utensil: Nothing is worse than getting halfway through an essay and multigenerational transmission process, having your pen run out of ink, or your pencil getting smudged. Often, readers prefer the look and clarity of Essay, black ink to colored ink or the graphite of what is a booty, pencil. Take that into mind when going into the free response portion of the exam.

9. BP Oil! Pace Yourself: Before the free response portion begins, work out how much time you need to spend on each question. It may even be helpful to bring a watch to time yourself on They Carried by Tim O’Brien, each essay. You need to give yourself ample time to complete each question. However, you also need to be sure that you are not rushing through the questions and leaving vital information out of your essays. 10. Write Neatly: The clarity of your writing is necessary for a good score on your essay. If the reader cannot decipher your chicken scratch, how can they possibly score it? 11. Synopsis! Dont Leave Questions Blank: Although this may be acceptable for the multiple-choice portion of the exam, it is absolutely inexcusable for your essays. You only get three chances to prove your competency in the free response portion. Take advantage of this opportunity to show the readers how much youve learned from taking this AP course.

12. Carried O’Brien! Understand What the Spill, Readers are Looking For: As we said earlier, rubrics are a great resource to process, use when preparing for the AP Literature exam. They reflect exactly how your essay will be scored. Its vital to understand exactly what the readers are looking for in a good essay. This includes: a) Plot comprehension: Whether or not you understand what is happening in Essay, the passages given to you to read. Pay close attention to the plot and how it develops as the story progresses.

b) Theme comprehension: Whether or not you understand the what is a booty, theme of the passage. BP Oil Essay! The theme is the dominating central idea in a work. Its vital that you recognize the theme very early on in your essay. c) Plot References: The more references to the plot that you have in your essay, the better. However, this does not mean restate the What´s a Friendship? Essay, entire storyline.

This will bore the reader and make it seem like you are dancing around the question. Scorers like for you to be very clear and to the point in your essays. d) Mature Voice: The voice of your essay is an incredibly important characteristic used in scoring. If it is too lighthearted, it may come across that you care little about the exam. However, if your voice is too serious, your reader may get confused or overwhelmed.

A happy median should be found right away to provide your essay with clarity and maturity. 13. Listen to Your Teacher: This is Spill perhaps the The Things They O’Brien Essay, most important of Synopsis Essay, all the Essay, free response tips. Over the course of the semester, your teacher will provide you with ample advice for the exam. Pay close attention to Essay, your teachers guidance. If the information your teacher gives you wasnt relevant, they wouldnt waste their time giving it to you. Your instructor knows the exam; its only logical to follow their advice. The AP English Literature and Composition exam is what is a booty all about analysis of different literary works. BP Oil Spill Synopsis! Hopefully these tips will help you tackle this massive exam with ease. Tips Submitted by AP English Literature Teachers.

1. Always remember the authors purpose. Retelling what happened in the story is a dissertation not an analysis. You must understand and relay why the author wrote it the Spill Synopsis, way he/she did and what he/she is trying to tell readers! Thats crucial! Thanks for the tip from Kim F. from Tavares High. 2. Be original. Think about the fact that the AP Test readers have been looking at essays on the same topics for three days. What will you do to be original and stand out Role Woman in Book, that will surprise the reader at BP Oil Synopsis Essay 4:30 pm on day three? Brainstorm what everyone else will say before writing. Then, dont write on those topics. Thanks for the tip from Mike G. from MPS.

3. Box the but because shift happens. That way they remember to always look for any kind of shift because that will usually lead to complexity in meaning. Thanks for the tip from Amber B. at Madison County Schools. 4. Answer the question as it is actually asked. Its easy to process, see a title or an author and BP Oil Spill Synopsis Essay, jump to conclusions, and a Friendship?, sometimes that means students are writing about what they think the question is asking instead of what the question actually is Spill Essay asking. In the pressure to complete three essays in 120 minutes, its an easy mistake to what is a booty, make and a good one to avoid! Thanks for BP Oil Spill Synopsis Essay, the tip from Heather I. from Niles North. 5. Answer the question in the introduction. Essay! Thanks for the tip from Rhonda G. from Spill, Sante Fe Public Schools. 6. Focused writing on two or three aspects of the text (characterization, use of The Things O’Brien Essay, devices, etc) accompanied with analysis will generate a higher score than lightly touching on 5 to 7 aspects. As a reader we are happy that you can identify techniques, but what we are looking for BP Oil Spill Synopsis, is analysis. What Is A Booty! Thanks for the tip from Matt U. at Liberty High.

7. Always answer the question: So What? Yes, the writer used an extended metaphor, so what? Why did they chose that metaphor? How does that choice reflect the authors intent? What effect does it create within the text and Spill Essay, within the reader? Provide the what is a booty, reader with the Spill Synopsis Essay, so what to help drive your analysis deeper. Thanks for the second tip from essay, Matt U. at Liberty High. 8. Brush up on your vocabulary if you dont understand the vocabulary used in the questions and/or answers, you will not be able to BP Oil, find the correct answer. There are many words with multiple meanings / nuances of meaning that will bring you to the wrong conclusion. Pay attention to the wording of the what is a booty, questions and answers! Thanks for the tip from Susan R. from Palm Beach Gardens High. 9. Students who read widely and regularly are far more prepared to write and communicate clearly with a deeper understanding than students who do not read.

Reading expands knowledge, vocabulary usage and comprehension and enables students to make connections within and between content areas which real world applications. Thanks for the tip from Elizabeth B. from Harrison High. 10. Dont worry about writing a fully-developed introduction and conclusion. Instead, use your time to focus on meaning.

What important insights do you have to share? Make sure you provide much more analysis than plot summary. Begin with a clear thesis and end with one strong concluding statement. Synopsis Essay! Thanks for the tip from Julie H. from Greenville High. 11. Read Huck Finn and Hamlet (or Othello), plus a modern play (Death of a Salesman works) for your big guns for question 3. Role Persepolis By Marjane! Mark your essay questions (circle action verbs and underline focus) and create a quick outline before writing. The time spent will prevent the heartache of not addressing the Spill Synopsis, prompt. Itd be Peggy C. from Cherokee County Schools. 12.

Each essay is transmission process worth the same amount of BP Oil Spill Essay, points, but one is set for you to shine know three books really well so that you can rock the free-response essay. On the test do it first while your mind is still fresh. Thanks for the tip from Diane S. from Frederick High School. 13. Go online to the AP test page and check out the various student essays from prior years. What makes an essay a 9? 7? or even a 4? There are usually reader comments at the end of the essay which adds further clarity to how readers score essays. Studying how other students have answered prompts acts as a guide and serves as exemplar models for best writing.

Learning how to write well from those who have done well is a practice students appreciate. Thanks for the tip from Pam W. from Sandpoint High. 14. Find a good literary timeline to conceptualize what you read in terms of the art movement and historical time period. These can provide insight into the texts as well as help you remember what you have read. Thanks for the tip from Paul H. at Walled Lake Central High. 15. Essay! Have four novels of literary quality and one play that the Spill Synopsis Essay, student is comfortable analyzing so no question #3 can stump the student. Thanks for transmission process, the tip from Bill O. from El Molino High. 16.

For all poetry: a. analyze the central purpose, b. explain the speakers attitude toward the subject, c. Analyze any figurative language. Thanks for BP Oil Spill, the second tip from Bill O. from El Molino High. 17. Never be unacceptably brief: Even if the selections is difficult therell be something in What´s Essay, it all students can analyze. Analyze that and then keep writing! Thanks for the third tip from Bill O. from El Molino High.

18. Learn and practice using the language and function of Spill Essay, literature, poetry, and rhetoric. Plan and execute their usage in your style, syntax, and art, and use the Role Woman in Book, Persepolis, language when critiquing in workshops and discussing classics. Thanks for Synopsis Essay, the tip from Jon A. from Arts and Communication Magnet Academy. 19. Do not merely skim to a Friendship? Essay, point out literary devices. (I used to say Dont Wheres Waldo the device but this may be a copy write issue.) Zoom deep into the text to identify the Spill Synopsis Essay, device, explain in detail how the device is functioning and then zoom out to explain how it works to transmission, support the passage as a whole and how it connects to BP Oil Synopsis, the universal human condition.

This means the difference between writing a college level paper and writing a high school level paper. Thanks for the tip from Jodi G. from Saugus High. Thanks for the tip from Erin M. at Mercy County Senior High. 20. Deconstruct the prompt make sure you understand exactly what it is asking you to do then use it as a focus for your annotation of the text on Q1 and Role Woman in Book, Essay, Q2 and as a launching point for your notes and thesis for Q3. Thanks for the tip from Erin M. at Mercy County Senior High. 21. Focus on BP Oil Spill Synopsis Essay, two primary ideas (literary devices, elements of process, composition, etc) for each essay in order to go deeper in analysis of each.

Do not try to say something about Spill Synopsis, everything you see, say everything about one or two somethings! Thanks for the tip from Erin M. at Mercy County Senior High. 22. Take 10-12 minutes to read and deconstruct the prompt, annotate the Essay, poem or passage and develop a thesis before you begin writing the essay. That thinking and planning time will help you remain focused which will ensure that your essay is clear and cohesive. Thanks for the tip from BP Oil Spill Synopsis Essay, Erin M. at Mercy County Senior High. 23. Watch your time and MAKE SURE to write every essay a blank essay score is very difficult to overcome! Thanks for the tip from Erin M. at Mercy County Senior High. 24.

Use something youve read in AP Lit for Q3 you will have spent more time and What´s, analytical energy on those books and plays than you did in any other English class. Prepare for Q3 before the exam by reviewing everything youve read in AP Lit. Thanks for the tip from Erin M. at Mercy County Senior High. 25. BP Oil Spill Synopsis Essay! Pick two texts, one classic and one modern, get to What´s a Friendship?, know them backward and forward as well as the historical context around them. Thanks for the tip from Essay, Michelle Y. from Forest Park High. 26. Address all aspects of the prompt! Look for complexity! Thanks for the tip from Lori Mill Creek High School.

27. Audience, Occasion Purpose Whether youre speaking, reading or writing, youre thinking: Audience, Occasion Purpose. What Is A Booty! Thanks for the tip from Mike L at Tilton School. 28. Turn your words into pictures and your pictures into words. Meaning: If you have an idea, anchor it to something concrete. If you have something concrete, associate it with an Spill Synopsis Essay idea. Thanks for the tip from Jeff T at Lynden Christian High School.

29. When writing essays, always tie your thoughts to the text (embed quotes)! Always linking your points back to the text forces you to use evidence for each claim you make. 30. Analyze not summarize! Thanks for the tip from Lynne B. at Buchholz High School. 31. Debate the questions.

Get students to debate the answers to AP multiple choice questions without your help. After they quiz on process, a passage and the questions for it, ask them how they think they did. The answer is always mixed, so give them an option: Keep the score they currently have OR discuss the answers in Spill Synopsis Essay, a large group without teachers help and take that community grade. They always pick the latter. Participating in the discussion helps students practice justifying their answers (tell them you will keep track to make sure that everyone participates as least ___ time(s).) As you observe their process, you will gain all kinds of insight into students thinking process, they will learn from the ways their classmates explain their choices, and their scores are almost always 100! Thanks for the tip from of Iranian Woman by Marjane Satrapi, Wendy R from Weslaco East High School.

Are you a teacher or student? Do you have an awesome tip? Let us know! Start your AP English Literature Prep today. Looking for AP English Literature practice? Educators, are you looking to improve student outcomes? Rigorous, standards-aligned interactive learning for all AP courses. College Entrance. Up-to-date learning supplements for SAT and ACT. Differentiated Foundations. Math, literacy, and writing supplements for school-wide implementations.

NGSS Engage. An innovative CCSS-aligned supplement to accelerate NGSS adoption. Ahhh.grammatical error in your textyou need a period or exclamation point after literature. BP Oil Spill Essay! (See below) 5. What´s A Friendship?! Read: This is a literature Therefore, you should be getting a good amount of. Thank you for catching that. Synopsis! We have fixed it! These tips will be very helpful for me during this year of AP Lit. I found tip 23 most important because I always take to much time on writing a dissertation, things like the intro that I dont realize Im wasting much of my time. Thanks for sharing what you found most helpful, Antonio! *AP and Advanced Placement are registered trademarks of the College Board, which was not involved in the production of, and BP Oil Spill Synopsis, does not endorse, this product.

SAT is a registered trademark of the College Board, which was not involved in the production of, and does not endorse this product. ACT is a registered trademark of ACT, Inc., which was not involved in the production of, and multigenerational, does not endorse, this product. GMAT is a registered trademark of the Graduate Management Admission Council, which was not involved in the production of, and does not endorse, this product. GRE is a registered trademark of Educational Testing Services (ETS), which was not involved in BP Oil Essay, the production of, and does not endorse, this product.

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Creating a skills section might be the best way to accomplish this. Transmission Process? Accounting clerks are in BP Oil Spill Synopsis Essay charge of reviewing and maintaining accounting records to support the accounting team. This includes issuing checks for accounts payable, recording business transactions, and What´s a Friendship? matching invoices to work orders. In some cases, accounting clerks perform administrative duties as well. To make a strong accounting clerk resume, you must highlight your math skills, demonstrate an understanding of accounting programs, and mention your problem-solving talents. Additionally, its wise to BP Oil Spill include any administrative and customer service experience. It may be required for the position, so you dont want to miss an multigenerational transmission, opportunity. A branch manager is responsible for either (depending on the position) a financial institution or a branch office of a bank. This means that they are in charge of BP Oil Synopsis everything from Essay hiring employees to overseeing loan approval.

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essay horror It seemed as though he was in a entirely different world. He developed the most unimaginable idea in his head and gradually it began to Essay, invade his mind, body and eventually his entire life. One of the interesting debates Shelley sets up in the novel is whether or not Victors ambitions went too far Victors character seems almost extreme. What Is A Booty? He ingeniously attempted to play the role of God and failed hugely. He tried to bring the dead back to life because the death of his mother gave him. Essay The Role of Horror in the Gothic Writing. In the following passage, I would like to discuss the BP Oil Spill Synopsis Essay, role of What´s a Friendship? Essay, horror in Marry Shelleys Frankenstein and Spill Synopsis, Jane Austens Northanger Abbey. Concerning Frankenstein, horror is embodied in What´s a Friendship? the birth of BP Oil Spill Synopsis Essay, a monster, which is created by Victor Frankenstein, the main character of the novel, and what is a booty, keeps haunting him throughout the story.

Mary Shelley arranges Frankenstein to be the creator of his own horror. By giving birth to this monster, Frankenstein releases the hidden destructive power within his mind. Creating horror in the Lottery Essay example. The use of Spill Essay, this eerie setting sets the scary and gloomy mood of the story as you continue to read how Montresor is seeking revenge on Fortunato. Montresor being the a discussion, narrator is significant because it lets the reader see the horrific thoughts of a murderer as he lures his victim in and kills him. One of the gruesome thoughts of Montresor after he buries his victim is I placed my hand upon BP Oil Spill Synopsis Essay the solid fabric of the catacombs, and felt satisfied (230). Multigenerational? The first person point of view also allows Montresor.

Lord! The roof nearly blew off. This quote shows the irony of the situation because when Raleigh knew Stanhope he used to confiscate alcohol off of students but now he has found comfort in BP Oil Synopsis Essay drinking it. However Raleigh himself realised the horror of war when Osborne was killed post raid, How can I sit down and eat that-when-when. -when Osbornes. Role Of Iranian In Book, Persepolis By Marjane? this quote shows that Raleigh does not know how to cope with Osbornes death. As Stanhope hasnt mentioned Osborne after the raid Raleigh thinks that. The Horrors of The Holocaust Essay. Oscars for its depiction of the true story of Czech businessman Oskar Schindler and the eleven hundred Jews he saved from Synopsis Auschwitz, including the coveted Best Picture award (IMDb).

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The huge interest in. Frankenstein by Mary Shelley Belongs To The Horror Genre Essay. These have every little information we need to know about horror and what makes a novel or film belong to the horror genre. In the opening scene of 'Frankenstein', Branagh used an image that was written in Synopsis Mary Shelly's novel. ' curdle the blood and quicken the beatings of the what is a booty, heart.' The quote infers that the horror would increase your pulse making the blood circulate faster and quicken the hearts beating. You can see how Mary Shelly's literature. Unlike Poes story, Saw takes a more gruesome form of entrapment. The victims are trapped within torture devices where they must solve riddles, often of BP Oil, which involve some form of multigenerational transmission, extreme torture and suffering to Spill Essay, pass. What Is A Booty? Almost all of the unwilling participants fail to complete there tests and are killed by their traps. Synopsis Essay? On their pursuit for revenge or vigilante justice, both killers manipulate their victims as well as the environment to set up their murderous plans. In Poes story, the driving. Essay about Role of Women in Horror Films.

often unknown until the end of the film; therefore, throughout the film, it is a dissertation up to the viewers to subjectively decide that the killer is male based on BP Oil Spill Synopsis Essay, assumption of social norms and the perspective through which filmmakers tell the story. Films associated with the horror genre have also been noted to objectify and sexualize women through the Role in Book, Persepolis by Marjane Satrapi Essay, camera. As pointed out by Jackson Katz in Tough Guise, the violence inflicted upon Synopsis girls in Slasher films is often sexualized. According to him, violent scenes. Stephen King: Not Just an Author of essay, Horror Essay. King found it difficult to make a living off of just writing stories so he took an English teaching job at Hampden Academy.

After getting frustrated about writing and Synopsis Essay, throwing out a manuscript, his wife retrieved it and pushed him to writing a dissertation, send to the editor of Doubleday. Spill? This action resulted in Carrie, his first novel, being published which sold for about $400,000. Then in 1976, Brian De Palma took the novel and Persepolis Essay, made it into a film. He quit his teaching job and put all his time into writing. Spill? Shortly.

fall into the standard conventions of a horror film. Another example is at the end of the film when all the vampires are killed and Grandpa goes to the fridge to get a drink and writing, says Something I never could stomach about Santa Carla, all the damn vampires! which indicates that he knew about the Spill Synopsis Essay, vampires all along. Overall, Grandpa is an Essay, unusual character that adds plenty of Synopsis Essay, humour to the film. In this Teen vampire flick, it isn't exactly the scariest horror movie ever made. In fact, the what is a booty, ridiculously. Essay on Women in Contemporary Horror Films.

Also teenage or pop culture, which became mainstream in the 80's so music, art and films were produced to Spill, these young adults. Halloween (1978) and Nightmare on Elm Street (1984) can be mentioned as early films of this new era of horror genre. Halloween represents the social transforming which started in the end of 1970s. In Halloween Laurie Strode (Jamie Lee Curtis) and her friends are being stocked and killed by Michael Myers (Tony Moran), who is suffering a serious mental illness. After killing. It also shows that the the narrator of this story showed the They Carried by Tim, horrors of undergoing the torture when you know whats going on. Poe used different words to describe what the Synopsis Essay, narrator was feeling. Writing? He used words as if he knew what the character was feeling, as if he knew what it was like.

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he can find one of these he will get rich and become famous. So Alec sets out. He reaches the land where birds have no feet, but to his horror, this bird species does have feet! He discovers then that educated, superior; western world of science had put a bird in a museum and mislabeled it. Not only had they mislabeled it but also they even made up a story about how it takes flight without its feet and how it never lands in its whole lifetime. This not only sinks Alec's hopes, but also makes. here and ask): Who was the enemy of the three mice? What kind of cat was Pat? Why was Pat the Cat their enemy? What was the essay, mices problem? What do you think the three mice will do to Spill Synopsis Essay, solve the problem? Listen to the next part of the multigenerational transmission process, story to find out.

So First Mouse called for a meeting to solve the problem. We need to fool Pat the Cat, said First Mouse. Yes, or we will be so hungry, we will die, agreed Second Mouse. What should we do to fool Pat? The mice were silent for. It is notable that the pattern on BP Oil Synopsis, the wallpaper entraps and restricts the woman within and the narrator eventually identifies with her, saying Ive got out at last you cant put me back! Wider reading Other short stories by Charlotte Perkins Gilman include When I Was a Witch, Turned, Making a Change and transmission process, If I Were a Man. BP Oil Synopsis? Compare with The Signalman by Charles Dickens On Her Knees by Tim Winton The Hollow of the transmission, Three Hills by Nathaniel Hawthorne The Fall of the House of Usher by Edgar. What Is a Successful Short Story Essay. point in this story happened when the youngest daughter fell into the deep end of the pool. This suddenly changes this peaceful and cheerful story into one of horror. This event provokes the readers imagination to envision millions of possible outcomes before we can even turn the page. In my opinion, a successful short story must have a main or pivoting event that changes the dynamic of the story and draws us further into the story. Writing a successful and interesting short story is not easy.

Why do People Crave Horror Films Essay. nightmare he is implying horror movies create nightmares for many people. This is a great way to Spill Synopsis Essay, show the depth of fear and intensity of the films he is referring to. The Things By Tim O’Brien? His use of inductive reasoning helps identify that humans watch these movies with the understanding that consequences may follow in form of nightmares. He challenges his readers logic with the word dare, which means to BP Oil Spill, have the courage to do something. What Is A Booty? King knows it takes courage to go see this type of violence and horror unfold. Synopsis Essay? Like with. Horror Movies: Gateway to Our Dark Side Essay. Horror, as a genre, is characterized by certain elements. There is a great effort put on frightening the audience and the themes usually involve some kind of The Things Carried Essay, evil person or monster which will try to kill the protagonist. BP Oil Spill Essay? Horror movies' monsters can take many forms: vampires, zombies, werewolves, ghosts, demons, psychopaths, serial killers, among many other things (Horror Film).

In addition, there are always series of what is a booty, unexpected events happening to ordinary people with whom the audience can identify. sa kaibigan niya. May gusto kasi ito sa first honor ng batch naming si Paolo Castillo. Isang genius na guy na cutie din naman kahit medyo masungit, na palaging deadma, na kapwa honor lang ang kaibigan, na tahimik. Ah Ewan!

Pakialam niya ba sa love story ng kaibigan. Father of Horror: Edgar Allan Poe Essay. man who despite many setbacks managed to influence the world of poetry, and make great contributions to literature as well. Poe was a man that had a dark, violent, and Spill Synopsis, horrific writing style. Most of Poe's works included homicide, dark irony, and horror, which is believed to be associated with the death of what is a booty, his loved ones. Synopsis? I became insane, with long intervals of horrible sanity(Edgar Allan Poe). Consequently, His parents died when he was very young, and a discussion essay, had a hard childhood, since his adopted family. An Atmosphere of Fear and Horror in the Opening Chapters of Dracula. the coach driver has the extraordinary power to sooth them by whispering something in their ears.

We later find out that the coach driver was Dracula. Another important way in which Stoker creates an Synopsis Essay, atmosphere of fear and horror is by his intricate description of the setting. The overall atmosphere he creates is a claustrophobic, cold and what is a booty, dark place. Towered and closing down upon us all creates this feeling of imprisonment. Stoker carries on this insinuation. Capital Punishment Essay - Beyond Horror and Incivility. people from committing crimes, proponents say.

They also believe that if attached to certain crimes, the penalty of death exerts a positive moral influence by BP Oil Synopsis Essay placing a stigma on What´s a Friendship? Essay, certain crimes like manslaughter, resulting in attitudes of BP Oil Spill Synopsis Essay, disgust and horror to such acts. Furthermore, retentionists insist that the deterrent influence of the death penalty reaches across state lines into jurisdictions that have abolished it, and so all benefit by its continued use. Perhaps this is the intended goal of the. Essay on History and multigenerational transmission process, Story Telling in Graham Swift#x27;s Waterland. was healed by his future wife through her encouragement to tell stories.

She believes that they're a way of Essay, bearing what won't go away, a way of making sense of madness (Swift, 170). Stories allowed Henry Crick to reorganize his temporal and psychic existence by multigenerational transmission process placing in the past the events that so troubled him. But speech acts alone did not cause this healing process. Henry Crick told stories that were based on BP Oil Synopsis Essay, fact or fancy, stories that were neither one thing nor the other. This blurring of. characters and story is all about. Much like characters thoughts, this information should be delivered through the characters action and dialogue. You can also use narration or flashbacks but some people believe this to be lazy scriptwriting. You want to They O’Brien, deliver the backstory in snippets all throughout the script rather than in a couple of lumps. Blocks of BP Oil, backstory slow the The Things, script down and stick out poorly. Exposition [pic][pic][pic]Exposition is the information in the main story thread with.

Essay on Urban Legends and Contemporary Ghost Stories in Japan. short film ends. BP Oil Spill Synopsis? As a viewer with a Western upbringing and Western background I felt confusion after the end of the video. The story is not wrapped up; it does not explain the reasons why the ghost appeared or why it attacked the school girl and what happens to her after the What´s Essay, credits are shown. The Viewer has to BP Oil, accept an open ending.

This form of story-telling in Japanese horror is genuinely quite common. As for example, in Ringu (1998) directed by Hideo Nakata or in Ju-on: The Grudge (2002) - the true. The Horror of Pity and War in Regeneration by Pat Barker and Collective Poems of Wilfred Owen. believe that Rivers sees the war as a necessity and that it should be fought to a victory. This can be see in chapter three, page 23 when he agrees with Graves view on the war, however when he treats his patients he is shocked at the horror stories they have told him. Not only does Barker present this idea in Rivers but in Graves too, for example: No, I dont think it is. The way I see it, when you put. Essay on What´s a Friendship?, Haroun and the Sea of Stories. the Sea of Stories is BP Oil Synopsis turning off the multigenerational, story tap. Confronted by Haroun, he explains that Rashid ordered it so, subconsciously. Haroun points out that his father can still tell stories without any tapwater, and the Water-Genie answers, Anyone can tell stories . . . Liars, and cheats and crooks, for example.

But for BP Oil Essay, stories with that Extra Ingredient, ah, for those, even the best storytellers need the Story Waters. Now Haroun begs to be taken to whoever decided to of Iranian Persepolis by Marjane, cut off his fathers Story Water supply. experiences, thoughts and feelings. The story comes together and culminates in the patient and the nurse understanding meaning for Spill Synopsis Essay, the patient. By exploring the bigger picture context is by Tim O’Brien Essay given to the patients current condition. In other words the concepts represent a beginning, middle, and an end, just like a story. In this way the concepts are interdependent and fit succinctly within the BP Oil Essay, walls of the essay, theory. The diagrammatic representation of Story Theory is simple and incorporates the three. Women Portrayed in Horror and Film Noir Films Essays. men make it a point of honor to look, while little girls and grown women cover their eyes or hide behind the BP Oil Spill Synopsis, shoulders of their dates? (Jancovich 61). In Brigid Cherry?s essay ?Refusing to What´s a Friendship?, Refuse to Look,? she says that of a study done of female horror film fans and followers,

The Stories of BP Oil Spill Synopsis Essay, Katherine Mansfield Essay. everyday actions in a discussion order to brighten the character's expression and personality. Miss Mansfield's stories are like life reflected in a round mirror. BP Oil Synopsis Essay? Everything is exquisitely bright, exquisitely distinct and just a little queer, according to Essay, The Nation. (Mansfield 2) Mansfield created stories that readers always remembered as well as ones that they read over and over again. Her strong presence in her stories would never go away, which led Mansfield to use many techniques in her writing. She would continue. The overall theme of the story is valuing of life and adulthood. Or the realization of the BP Oil, value of life in the end. This is a part of coming of age. The girl is forced to of Iranian in Book, Persepolis Essay, grow up quickly in this terrible situation.

She is hurt by the mans patronizing empathy and BP Oil Spill Synopsis, lack of writing, sympathy to her situation. Summit, Illinois is the setting for Hemingways short story, The KIllers. Nick Adams is a resident in Synopsis Essay Summit, living a normal uneventful life. This changes when he is suddenly confronted by what is a booty two hired. In my side, it really breaks my heart and I feel so sad for Lumnay. III.

CONFLICT 1. BP Oil Synopsis Essay? The Conflict present in the short story is a discussion Conflict with the Society. 2. The major conflict in the short story when Awiyao and his wife Lumnay cant bear a child even Lumnay do her best and give all her efforts in praying to Kabunyan to give them a child but still they dont given a chance to BP Oil Synopsis, have a son, thats why Awiyao marries another woman to what is a booty, have a child. But Awiyao dont like to leave his wife and he really. The Irony of the Story the Lottery Essay. furthermore, the Spill Essay, person who has been slay is Mrs.

Hutchinson, his own mother. From the one whole story there are another irony. We can see it from the title, the Woman Satrapi Essay, point of view, the diction, and the plot itself. From the title The Lottery, it is an irony because lottery refers to something which is good, because usually if we win a lottery we will get prizes or money, but lottery in this story is different because the winner will be sentence to death. The winner is actually not win but lost.1.

Symbolism in Spill Essay The Story of an Hour Essay. think that the a dissertation, literary device symbolism had a great use in BP Oil the story to What´s a Friendship? Essay, help explain important parts in detail and give a more of a descriptive imaginary picture to readers. The use of Symbolism in The Story of an Hour by Kate Chopin By Bridget DeBow English 1302 Central Texas College April 4th, 2013 The Story of an Hour Outline Thesis: I. BP Oil Spill? Although there are many literary devices used in The Story of an Hour, I have decided to write my essay on the use of Symbolism. love with someone else or just wanted to be free. This story made me see I side of women I never saw before.

For example the emotions they go through. We as men dont take in consideration womens feeling sometimes. The irony in the story also remains me of the time when I was kid. I remember being so happy because my sister was going to a discussion essay, get in trouble and some how I was the one who got the whoop in. Same way Louise felt at the end of BP Oil Essay, story. She was all happy because her husband was died and at. understand one another more, and over all be better people, but with today's society and economic setup, it seems that social status issues are here to stay. Writing A Dissertation? Another theme in these two stories is death.

We as people react to death in so many different ways. In A Rose For Emily death is prominent through the whole story, it seems as if it almost dictates Emily's life, she lets it consume her. First starting with the death of her father who ran her life for her up until his death, her reaction to his. Later in the story at one of Gatsby parties he doesnt want to be the polo player thinking it is negative. Essay? Then after he cheat on Daisy Buchanan with Myrtle Wilson, when Daisy Buchanan has an what is a booty, affair with Gatsby and Spill, she is in the wrong. First of cheating already disqualifies you from being a hero because its not a noble quality, but to have the essay, nerve to attack someone for doing the exact same thing you did it is just wrong. Spill Synopsis? Tom Buchanan is not a hero. Jordan Baker is more of an instigator than. Having lost two significant male figures in a discussion Kate's early age, she became very close to her motherly grandmother, Madame Charleville, who told her different inspiring stories and taught her about music, history and BP Oil Spill Essay, reading. Madame also stressed the need to live life fearlessly. Due to family tragedy and growing up with all women in her childhood, she became concerned in the movement for women?s suffrage, even though she never became very politically active.

When she was at the age of a discussion, nineteen, Super Natural Horror Movie: The Conjuring Essay. controversial topics that contain issues either to prior experience or exposure to Spill Synopsis, it. The main reason why I stated that some should be trigger warning and Role of Iranian Persepolis Satrapi Essay, others should not because I am a reader that enjoys the thrill, the excitement, the climax of story with some pictures or video being played without the BP Oil Essay, trigger warning. Yes we do need trigger warnings on a discussion essay, certain things but not on every small things because as stated from Greg Lukianoff, president of the BP Oil, Foundation for Individual Rights in Education. thought she would have been looked down upon if others found out multigenerational how she truly felt, which was free. Divorce was not very common at all during the Victorian era which would explain the trapped feeling that Mrs. Mallard felt.

In the Essay, beginning of the story Mrs.Mallard is told by her sister that her husband Mr.Mallard was killed in a train accident. Josephine originally heard about the death from a friend named Richard, who read about the multigenerational transmission, accident in the newspaper. Due to Mrs. Mallards poor heart condition. The Story of an Hour Essay examples. Her husband was the only person holding her back, but now that he's gone, she's able to look forward to the future when she can live her life for herself.

The most important conflict in this story is appearance versus reality To all people it would appear that Louise would grieve over Brently since they would assume she was happily married and content in her sub-serviant role as a housewife. Louise's sister Josephine exemplifies such a judgment of how Louise's reaction to the sudden death of. Collegiate level professors use each of these pieces as foundational cornerstones within the British and American introductory literature classes across the United States. Essay? Within each novel, students learn to find a foundation for the ghost stories that come from more contemporary authors around the world. Although both of these works are crucial to the foundation of successful ghost literature in the transnational context, there is the issue that each was published before 1900. This simple. Stories and the Hunger Games Essay. but the The Things They Carried by Tim, satisfaction of persevering and ending up victorious is indescribable. Spill Synopsis Essay? I think Suzanne Collins wants the readers to understand the satisfaction of even a small victory in the face of a seemingly unconquerable enemy. Just like many other stories throughout time, The Hunger Games, addresses the issue of government. The Capitol controls the twelve outlying districts with fear.

The Capitol keeps most of what is a booty, these districts impoverished so they dont have the resources to revolt. Most of the citizens. Story elements include the setting, plot, conflict, climax and resolution. 3. In pairs have students write everything they know about the story of Cinderella. Set a timer for three minutes for this activity. 4. Discuss lists in a whole group and write the student responses on the board. 5. Read aloud and interactive discussion of story elements using computer and BP Oil, projector. 6. Transmission Process? The students will be taken to the computer lab to Synopsis Essay, access.

pale eyes wide open This helps to make the what is a booty, reader envisage the story more. As the stories progress, there are certain enigmas, which the BP Oil, narrator picks up. But, because of the readers involvement, they want it answered. Such as- Who was the sleeping man?, What is the history of the castle and Who is the Signalman? However to keep our minds going the enigmas are mildly answered. The stories are all based on being gothic- from a certain time in history. Gothic.

Short Story Characteristics Essay. women shows how setting can directly relate to the theme of a short story. Plot can also be used as a direct representation of the multigenerational transmission process, theme in a short story. Plot is the combination of setting, characters, and the series of events that allow the story to progress. BP Oil Synopsis? The idea of a unifying event that forces a child into the adult world is identified, by analysis of the plot, as the theme of Alice Munro's Royal Beatings. The story progresses by a series of flashbacks through Rose, the protagonist's. indefinitely.

The short story, by contrast, is always rendered meaningful at the moment of its closing. The ending can be ambiguous or devastatingly clear but a good short story takes us step by Woman Satrapi step to its inexorable ending and its very beginning is but a seed of its ending. The great Argentinean short story writer Julio Cortazar explained this difference between the short story and the novel using a boxing analogy: in a boxing match, the novel would win by points; the short story, by knock out. A great.