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Smalls-Mantey, Sample F31 Application and Summary Statement. We are truly indebted to the grantee who allowed us to essay focus post this outstanding F31 sample application to help the hbs essays 2015, next generation of investigators write applications. The F31 sample application below doesn’t reflect the complex layouts of the actual SF 424 PDF forms. Essay Focus Statement. Instead, we’ve provided a list of key fields from those forms and the applicant’s responses. In addition, we redacted some applicant-specific information such as contact information, publications in press, and budget specifics. The text of this application is copyrighted. You may use it only for nonprofit educational purposes provided the romeo, document remains unchanged and the PI, the focus, grantee organization, and NIAID are credited. Questions? Contact deaweb@niaid.nih.gov. We also offer other outstanding Sample Applications. [Our Web version of writing paper this sample application can't reproduce the layout of the actual PDF forms.

Here is the summary that eRA Commons generated at focus, the start of the sample app, which reflects many key fields from that form.] Project/Performance Site Locations. [Our Web version of essay this sample application can't reproduce the 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 essay focus, 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 jane elliott, 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 focus, most recent vaccine trials. However, this approach has not been successful in containing viral replication in vaccinees that have become HIV-infected. Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. In ADCC, Fc-bearing Abs bind viral epitopes coating an personalized preschool writing infected CD4+ target T cell and an Fc receptor bearing effector, most commonly natural killer cells (NKs), bind the focus statement, Ab and use perforin to deliver granzymes which induce apoptosis in hbs essays 2015, 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 essay, 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 essay, mAbs based on epitope specificity, affinity, potency, breadth, IgG isotype, and Fc type. We will also evaluate whether ADCC is disparate from classical neutralization. Finally, we will use microscopy to examine the synapse between effectors, Abs, and targets. The outcome of this research will provide insight into the characteristics of 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 essay focus statement mediate protection from hbs essays 2015, lentiviral infection. We hypothesize that variations in ADCC activity of sera are dictated by the amount, specificity, and focus statement subclass of HIV-specific antibodies. Aim 1 : Characterize the style referencing, potency of sera of HIV-infected individuals in ADCC. Aim 2 : Characterize the essay focus statement, specificity and breadth of antibodies with ADCC activity.

Aim 3 : Characterize the structure and function of the target-effector synapse using both fixed and hbs essays 2015 live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and 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 essay focus statement characterize the Abs that mediate ADCC and image the functional synapse formed by cellular components involved in ADCC with the goal of defining new goals for hbs essays 2015, the development of HIV vaccines and therapeutics. Gallo RC, Salahuddin SZ, Popovic M. Frequent detection and statement isolation of love essay cytopathic retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS. Essay Statement. Science. 1984; 224: 500-3.

Barre-Sinoussi F, Chermann JC, Rey F, Nugeyre MT, Chamaret S, Gruest J, et al. Isolation of romeo essay questions a T-lymphotropic retrovirus from a patient at risk for essay focus, acquired immune deficiency syndrome (AIDS). Style Referencing. 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 essay focus, antibody protection against HIV. Nature. Hbs Essays 2015. 2007; 449(7158): 101-4. www.hivresearh.org. RV144 Phase III HIV Vaccine Trial Press Release. 2009. Essay Statement. Chung A, Rollman E, Johansson S, Kent SJ, Stratov I. Love Essay. The utility of focus ADCC responses in HIV infection.

Current HIV research. 2008; 6(6): 515-9. Eye Of The Storm Jane Elliott. 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. Essay Focus. 1977; 118(3): 729-35. Thesis Recommendation For Future. Forthal DN, Landucci G. In vitro reduction of virus infectivity by essay, antibody-dependent cellmediated immunity. Journal of immunological methods. 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 HIV-infected subjects.

J Virol. 2008; 82(11): 5450-9. Hbs Essays 2015. Gomez-Roman VR, Florese RH, Patterson LJ, Peng B, Venzon D, Aldrich K, et al. A simplified method for the rapid fluorometric assessment of antibody-dependent cell-mediated cytotoxicity. Journal of immunological methods.

2006; 308(1-2): 53-67. Migueles SA, Osborne CM, Royce C, Compton AA, Joshi RP, Weeks KA, et al. Lytic granule loading of CD8+ T cells is required for essay statement, HIV-infected cell elimination associated with immune control. Immunity. 2008; 29(6): 1009-21. Vago L, Perna SK, Zanussi M, Mazzi B, Barlassina C, Stanghellini MT, et al. Loss of mismatched HLA in leukemia after stem-cell transplantation. N Engl J Med. 2009; 361(5): 478- 88. Ziegner U, Campbell D, Weinhold K, Frank I, Rutstein R, Starr SE. Deficient antibodydependent cellular cytotoxicity against human immunodeficiency virus (HIV)-expressing target cells in perinatal HIV infection.

Clin Diagn Lab Immunol. 1999; 6(5): 718-24. Brunetta E, Fogli M, Varchetta S, Bozzo L, Hudspeth KL, Marcenaro E, et al. The decreased expression of Siglec-7 represents an early marker of dysfunctional natural killer cell subsets associated with high levels of HIV-1 viremia. Blood. 2009.

Ljunggren HG, Ohlen C, Hoglund P, Yamasaki T, Klein G, Karre K. Afferent and in essay 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. Essay. Expression of HLA class I-specific inhibitory natural killer cell receptors in HIV-specific cytolytic T lymphocytes: impairment of specific cytolytic functions. Proceedings of the preschool writing, National Academy of Sciences of the United States of 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. 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 focus, HIV-1 elite controllers. AIDS. For Future. 2009; 23(8): 897-906. Focus. Dalgleish A, Sinclair A, Steel M, Beatson D, Ludlam C, Habeshaw J. Romeo And Juliet Essay Questions. Failure of ADCC to 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. Antigenic specificity of antibody-dependent cell-mediated cytotoxicity directed against human immunodeficiency virus in antibody-positive sera. J Virol. 1989; 63(2): 584-90. Doria-Rose NA, Connors M. Antibody-secreting B cells in HIV infection. Curr Opin HIV AIDS. 2009; 4(5): 426-30. Burke B, Barnett SW. Broadening our view of protective antibody responses against HIV.

Current HIV research. 2007; 5(6): 625-41. Sattentau Q. Avoiding the void: cell-to-cell spread of focus statement human viruses. Nat Rev Microbiol. For Future. 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. 1994; 153(9): 4268-80.

Jolly C, Kashefi K, Hollinshead M, Sattentau QJ. HIV-1 cell to cell transfer across an Env-induced, actin-dependent synapse. J Exp Med. Essay Statement. 2004; 199(2): 283-93. Groot F, Welsch S, Sattentau QJ. Efficient HIV-1 transmission from eye of jane, 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 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 focus, two distinctive perspectives in my graduate career, something most PhD scientist don’t experience until their first post-doctoral experience. While at 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. 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 preschool writing, FAES at NIH I have access to courses to improve lab specific skills and foundation knowledge.

I have already participated a Vaccine Development course. At Oxford I will take seminars at the Centre for essay focus, Excellence in Teaching and 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 scholarly research. Physical Resources : Both labs have sufficient funding to hbs essays 2015 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 focus statement, lab. Safety – In order to ensure our safety from hazardous blood samples we work under laminar flow hoods in a P2 facility with P3 practices. Samples – We receive blood samples from patients enrolled within our protocol. Romeo Act 4. We also have access to the NIH blood blank for normal human blood samples. As provided by Quentin Sattentau: “Oxford University is held in statement, high international regard for its strong teaching and research background and hbs essays 2015 its unique and personal teaching style.

Within the 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. Essay Statement. The Department has a strong graduate support structure, in which each student has a principal supervisor, the love definition, possibility of a cosupervisor, a departmental advisor, and a college advisor. The supervisor(s) will generally meet with the student multiple times a term, and each of the statement, non-supervisory mentors will meet with the candidate approximately once a term and will file reports on the candidate’s progress. The candidate will be expected to attend a series of high profile external seminars that take place once a week in essay, term time, and a regular series of internal seminars from focus, leading and more junior University and Departmental investigators and students. Adjoa will attend and personalized preschool writing paper participate in the weekly lab journal club, in essay focus, which scientific papers are taken apart critically by members of the eye of the storm jane essay, group, and essay focus 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 thesis about recommendation for future other reading and writing. Adjoa will have her own writing up space close to focus statement her work bench, equipped with a portable computer and if required, an external screen. Adjoa will have full access to all core facilities, including the imaging suite with multiple light microscopes, a flow cytometry and sorting suite, an electron microscopy facility with full technical and scientific support. About Recommendation For Future. We also have strong connections with the biochemistry and focus chemistry departments and thesis recommendation the Weatherall Institute of Molecular Medicine at essay, 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 elliott ancillary equipment, in which HIV-1 is grown and used to infect cells.

Flow cytometry. Focus. 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 hbs essays 2015, molecular biology carried out under containment levels –I, -II and –III. Light microscopy. Focus. The department has 5 confocal microscopes of varying sophistication and definition essay resolution, two of essay which are set up for romeo and juliet essay, live cell immunofluorescence work. The department has three recently acquired high-speed CCD cameras and statement associated microscopes for live cell immunofluorescence analysis. The Sattentau laboratory has a Zeiss Pascal confocal microscope and jane elliott a new CCD camera set up with a Zeiss Axiovert 200 microscope for fixed and live cell work in essay focus statement, 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 Oxford (Kay Grunewald) is located at the University Churchill campus, which is 4 miles from the Pathology Department. He maintains and 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 this sample application can't reproduce the layout of the actual PDF forms. See below for the biographical sketch attachments.]

My interest in studying infectious diseases stems from the 2001 anthrax scare in the Washington D.C. area. I already had a strong interest in science but through my participation in the Gene Search Science Project at Catholic University the hbs essays 2015, following summer I came to understand how medical research impacts treatment for routine illnesses and also plays a role in essay focus statement, national defense. Since middle school I have been preparing for a career in science. At Eleanor Roosevelt High School I was enrolled in the rigorous Science and Technology Program during which time in participated in the Gene Search Project and researched biochemical pathways that influence sickle cell anemia with Dr. William Winters at Howard University Hospital. This experience taught me how research and medicine are intertwined and inspired me to become a physicianscientist. At UMBC I was a member of the Meyerhoff Scholars Program and an HHMI Undergraduate Research Scholar where numerous lab experiences, seminars, rigorous classes, and multiple opportunities for harvard style in essay, scientific presentations prepared me for focus, a career as a scientist. My lab skills and harvard in essay understanding of a research career increased exponentially during my four years in Michael Summers’ lab. Not only essay focus, did I learn about harvard style referencing protein purification, gel electrophoresis, NMR, and ITC while studying MuLV RNA packaging, but more importantly of the time and discipline necessary for excellent research and the internal drive that keeps one focused. Working with Dr. Essay Statement. Nikola Pavletich I learned the hbs essays 2015, value of immediately organizing my data so that others could analyze my results quickly.

In the lab of Tom Cech I had the responsibility of designing a new protocol to statement 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 I’ve received at Columbia has given me a unique perspective with which I approach scientific questions. Under the tutelage of Mark Connors and Quentin Sattentau, leaders in their fields on nonprogressive HIV infection and viral transmission, my understanding of infectious disease and my capacity to pursue further research in these fields will be greatly enhanced. Hbs Essays 2015. In addition, the courses, medical preceptorships, presentations, teaching, and essay focus statement career development activities I am undertaking in about recommendation for future, the forthcoming years make me an 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. Focus. 2005. Composition and sequence-dependent binding of RNA to the nucleocapsid protein of eye of the storm elliott essay Moloney murine leukemia virus. Essay. Biochemistry. Personalized Preschool Writing. 44(10), 3735-3744. Essay. 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. Hbs Essays 2015. 2010. Structure of a conserved retroviral RNA packaging element by NMR spectroscopy and cryo-electron tomography. J Mol Biol.

404(5), 751-772. Smalls-Mantey A, Winter W. 2003. Biosynthesis of statement 2,3-diphosphoglycerate in preschool paper, Erythrocytes. Focus. Poster presentation at ERHS Science Fair and Prince Georges County Science Fair. Hbs Essays 2015. March 2003.

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

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

Poster presented at 2007 UMBC Undergraduate Research and Creative Achievement Day. April 2007. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. 2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells. Poster presented at 2010 Keystone HIV Vaccines (X5) Meeting. March 2010. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu.

2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells. Poster presented at 2010 Oxford Pathology Department Graduate Student Symposium. June 2010. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. 2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells.

Poster presented at 2010 NIH/Oxford/Cambridge Scholars Research Colloquium. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. 2010. Essay Statement. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells. Poster presented at 2010 International AIDS Vaccine Conference. September 2010. Smalls-Mantey A. 2010.

Antibody-Dependent Cellular Cytotoxicity and the Control of HIV. Essay. 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 considered one of the very top priorities in focus, the field of HIV-specific immunity. Our laboratory has assembled the best techniques and reagents in the field to carry out writing this project. Focus. We have extensive experience in HIV-specific antibodies and personalized preschool paper cytotoxicity. In addition we have a long track record of training students to prepare them for focus statement, a carreer in personalized writing paper, 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 focus statement, 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 referencing in essay 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 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. Public Health Service, Medical Officer, National Institute of essay Allergy and Infectious Diseases, Clinical and Molecular Retrovirology Section, Laboratory of Immunoregulation, National Institutes of Health, Bethesda, MD December 2007- Present Chief, HIV-Specific Immunity Section, Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD. Recent Honors and Editorial and Review Board Activities. 2009 - U.S.

Public Health Service Outstanding Service Medal 2009 - present Vaccine Production Development Team Scientific Advisory Board, University of Pennsylvania 2009 - present HIV Enterprise Host Genetics and HIV Diversity Working Group 2010 - present Editorial Board, Journal of Virology. C. Peer-Reviewed Publications. Migueles SA, Sabbaghian MS, Shupert WL, Bettinotti MP, Marincola FM, Schwartz D, Sullivan J, and romeo act 4 essay questions Connors M. HLA B*5701 is highly associated with restriction of 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 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 restrict HIV replication is not caused by loss of recognition of autologous viral gag sequences. J Virol 2003; 77(12); 6889-6898. Iyasere C, Tilton JC, Johnson AJ, Younes S, Yassine-Diab B, Sekaly RP, Kwok WW, Migueles SA, Laborico AC, Shupert WL, Hallahan CW, Davey RT Jr., Dybul M, Vogel S, Metcalf J, and Connors M. Diminished proliferation of HIV-specific CD4+ T Cells is associated with diminished IL-2 production and preschool paper recovered by exogenous IL-2. J Virol 2003; 77(20); 10900-10909. Tilton JC, Luskin MR, Johnson AJ, Manion M, Hallahan CW, Metcalf JA, McLaughlin M, Davey RT Jr., and Connors M. Essay Focus. Changes in paracrine IL-2 requirement, CCR7 expression, frequency, and cytokine secretion, of HIV-specific CD4+ T cells are a consequence of antigen load. J Virol 2007; 81;2713-2725. Migueles SA, Osborne CM, Royce C, Compton AA, Joshi RP, Weeks KA, Rood JE, Berkley AM, Sacha JB, Cogliano-Shutta NA, Lloyd M, Roby G, Kwan R, McLaughlin M, Stallings S, Rehm C, O’Shea MA, Mican J, Packard B, Komoriya A, Palmer S, Wiegand AP, Maldarelli F, Coffin JM, Mellors JW, Hallahan CW, Follman DA and Connors M. Lytic Granule Loading of love definition CD8+ T-Cells is essay statement Required for HIV-Infected Cell Elimination Associated with Immune Control. Immunity 2008; 29 (6): 1009-1021.

Migueles SA, Weeks KA, Nou E, Berkley AM, Rood JE, Osborne CM, Hallahan CW, Cogliano-Shutta NA, Metcalf JA, McLaughlin M, Kwan R, Mican JM, Davey RT Jr, Connors M. Defective human immunodeficiency virus-specific CD8+ T-cell polyfunctionality, proliferation, and eye of the storm jane elliott essay cytotoxicity are not restored by antiretroviral therapy. J Virol 2009 83(22): 11876-11889. Doria-Rose NA, Klein R, Daniels M, O'Dell S, Nason M, Lapedes A, Bhattacharya T, Migueles S, Wyatt RT, Korber BT, Mascola JR, Connors M. Breadth of HIV-Specific neutralizing activity in sera: clustering analysis and association with clinical variables. J Virol 2009 84(3): 1631-1636. Essay Focus Statement. 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 eye of elliott Mascola JR. Rational design of envelope identifies broadly neutralizing human monoclonal antibodies to HIV-1.

Science 2010 329:856-861. Essay Statement. 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 the storm, HIV-Specific Immunity Section of the Laboratory of Immunoregulation is focused upon mechanisms of immunity to HIV. The laboratory is staffed by PhD and MD scientists with extensive experience in the humoral or cellular immune response to HIV. In addition we have close collaborations with investigators at the NIAID Vaccine Research Center in essay focus, the area of thesis about for future structure-function relationships of 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 the research budget of the Division of Intramural Research. Essay Focus. 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 d’Immunologie, Marseille, France 1999 - 2003 Reader, Section of Infectious Disease, Imperial College London, London UK 2003 - Professor of immunology, Department of Pathology, University of Oxford, Oxford UK. Other Experience and Professional Memberships. 1995 - Member, American Association for act 4 essay questions, the Advancement of focus statement Science 1998 - Member, American Society for definition essay, 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. Focus. 2005 - Member of the Vaccine Research Center (VRC) NIH Board of referencing Scientific Councilors 2007 - Member of BMGF Vaccine Monitoring Centre SAB 2008 - Member of NIH HIVRAD/Novartis Neutralizing Antibody Vaccine Consortium SAB 2009 - Member of statement study section for hbs essays 2015, Swedish Program for International Development 2010 - Member of NIH study section for the B cell Immunology Partnership Program for focus statement, HIV-1 Vaccine Discovery (U19) C. Selected Peer-Reviewed Publications. Most relevant to the current application. Martin N, Welsch S, Jolly C, Briggs JA, Vaux D and Sattentau QJ. (2010) Virological synapse-mediated spread of human immunodeficiency virus type-1 between T cells is sensitive to style referencing in essay entry inhibition. J. Virol. 84: 3516-3527. Essay Statement. Gonzalez N, Bermejo M, Calonge E, Jolly C, Arenzana-Seisdedos F, Pablos JL, Sattentau QJ and Alcami J. (2010) SDF-1/CXCL12 production by mature dendritic cells inhibits the propagation of X4-tropic HIV-1 isolates at the dendritic cell-T-cell infectious synapse.

J Virol. 84:4341-51 Groot F, Welsch S and hbs essays 2015 Sattentau QJ. (2008). Efficient HIV-1 transmission from macrophages to focus statement 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. 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. Style Referencing. 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. Focus Statement. Virol. 81: 13916-13921. Jolly C, Mitar I and Sattentau QJ (2007). Requirement for an intact actin and tubulin cytoskeleton for efficient HIV-1 assembly and spread.

J. Virol. 81: 5547-6 Jolly C and about for future Sattentau QJ. Human Immunodeficiency virus type-1 virological synapse formation in T cells requires lipid raft integrity. J. Virol. 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. Essay Statement. J. Exp Med. Eye Of The Storm Essay. 199: 283-193. Additional recent publications of important to 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 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 HIV-1 envelope glycoprotein antigenicity and immunogenicity. J. Mol. Biol. Aug 25th epub. Krashias G, Simon K, Wegmann F, Kok WL, Ho LP, Stevens D, Skehel J, Heeney JL, Moghaddam AE and 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 situ determined by cryo-electron tomography. Plos Pathogens August 2 (8): e83 Sheppard NC, Bates AC and Sattentau QJ (2007). A functional human IgM response to HIV-1 Env after immunization with NYVAC-HIV C. AIDS 19: 524-527. [Outside research support redacted from sample.] PHS Fellowship Supplemental Form, Research Plan. [Our Web version of this sample application can't reproduce the layout of the actual PDF forms. The attachments are included below. In the original version, these attachments met the page limits.]

Understanding the basis of an statement immune response that controls infection or provides sterilizing immunity remains a major goal in the search for eye of, 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 essay, containing viral replication in vaccinees that have become HIV-infected. Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. In ADCC, Fc-bearing Abs bind viral epitopes coating an infected CD4+ target T cell and an Fc receptor bearing effector, most commonly natural killer cells (NKs), bind the Ab and about use perforin to deliver granzymes which induce apoptosis in the target. Statement. We want to love study ADCC in infected patients to understand the magnitude and 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 elliott essay, our lab. Based on these findings, we will select the sera of essay statement patients with the hbs essays 2015, most ADCC, generate monoclonal Abs (mAbs), and characterize the mAbs based on epitope specificity, affinity, potency, breadth, IgG isotype, and focus statement Fc type.

We will also evaluate whether ADCC is disparate from hbs essays 2015, classical neutralization. Finally we will use microscopy to examine the synapse between effectors, Abs, and targets. The outcome of this research will provide insight into the characteristics of Abs that mediate ADCC that are likely important goals in the design of HIV vaccines or immunotherapies. Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is essay focus statement a function that has been shown to mediate protection from lentiviral infection. We hypothesize that variations in ADCC activity of sera are dictated by the amount, specificity, and subclass of HIV-specific antibodies. Aim 1 : Characterize the romeo act 4 essay questions, potency of sera of HIV-infected individuals in ADCC. In ADCC, Abs bind viral epitopes that are presented by infected CD4+ T cells. NKs expressing an Fc receptor bind the Fc domain of the essay focus statement, Ab and use perforin to deliver granzymes to 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 an HIV-infected CD4+ target T cell. 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 definition, serum of essay focus statement HIV+ individuals with various rates of progression and viral loads to determine which contain Abs capable of mediating the highest levels of ADCC. Compare the ADCC and neutralizing activity of patient sera. Aim 2 : Characterize the specificity and breadth of antibodies with ADCC activity. Our laboratory has panels of NAbs derived from questions, patients with known serum neutralizing or ADCC-mediating activity. Determine whether recognition of focus statement specific epitopes is required for and juliet act 4 questions, ADCC. Essay Focus Statement. Define the breadth of the polyclonal sera by eye of the storm jane, its ability to mediate ADCC in CD4+ T cells infected by different clades of HIV.

Titer serum total IgG, IgG1, and IgG3 binding infected CD4+ T cells. Aim 3 : Characterize the structure and statement 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 tomography, we will examine the synapse formed between NK and act 4 essay other cells with potential ADCC activity (macrophages and neutrophils) and infected target cells. We will specifically investigate: The structure of a functional ADCC synapse. The kinetics of ADCC function in real time and statement its relation to love definition essay antibody type and specificity. A role for antibody-dependent cell-mediated phagocytosis (ADCP) in elimination of HIV-infected cells. d. Receptors and effector molecules central to ADCC activity against HIV infected cells. Since being identified as the causative agent of AIDS in focus, 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 hbs essays 2015 prevent HIV infection but with the growing incidence of HIV infection and the limitations of current antiretroviral therapy, there is an focus statement imperative need for a vaccine.

One of the most critical areas of research on vaccines for HIV is the hbs essays 2015, search for correlates of immunity. At present the only method for determining the potential benefits of essay a vaccine with regard to eye of jane elliott essay protection from infection or lowering of statement viral load upon infection remains very large clinical efficacy trials. Determining correlates of immunity would tremendously accelerate the development process by permitting some estimation of clinical efficacy prior to a Phase III trial. Reliable correlates would permit screening of love vaccine candidates at a much earlier point in development with smaller numbers of participants. Over the focus, past 20 years, research into correlates of immunity to love essay HIV have largely focused on CD8+ cytotoxic T lymphocytes (CTLs) or neutralizing antibodies (NAbs). In addition to neutralization, antibodies (Abs) may fight viral infections by statement, antibody-dependent cell-mediated phagocytosis (ADCP) or by antibody-dependent cellular cytotoxicity (ADCC). In ADCC, Abs bind viral epitopes presented on hbs essays 2015, infected target cells and Fc? receptor-bearing effector cells such as natural killer cells (NKs) recognize and bind the essay focus, Fc? region of these Abs. Upon cross-linking of the Fc? receptor, NKs release perforin that punctures the target’s cell membrane and deliver proteases such as granzyme B (GrB) which induce apoptosis. ADCC has become a major focus within the field following two important studies highlighting its potential importance in providing immunity to HIV.

A recent study by Burton and colleagues demonstrated that neutralizing Abs with mutated Fc? receptor binding regions were less effective in protecting macaques from 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 style referencing, 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 essay statement showed that the hbs essays 2015, prime-boost combination of ALVACR HIV and AIDSVAXR B/E reduced HIV infection. This vaccine regimen was known to not elicit a strong cellular or NAb response. It does, however, elicit binding antibodies that have suggested to essay statement the field that the protective efficacy is likely mediated through ADCC 5. Our laboratory is part of romeo essay questions a large international effort to characterize ADCC antibodies from this trial in order to determine whether there were higher ADCC-mediating antibodies in uninfected vaccinees compared to those that were infected. Although HIV-specific ADCC is now considered an extremely important part of vaccineinduced immunity several basic questions regarding the statement, nature of an effective ADCC response have not been addressed. For example, the the storm essay, number, affinity, and specificities of essay statement 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 harvard referencing in essay, HIV-specific ADCC. Past studies of ADCC have been hampered by the availability of assays that are only partially quantitative. Essay Statement. There are four primary methods that people use to measure ADCC 6. The most prevalent is the style in essay, chromium release assays (CRA), which is highly variable, does not specifically account for the mechanism of killing, and is insufficiently quantitative 7. The ADCVI assay incubates targets, serum, and effectors for 24 hours and the supernatant is applied to the B95-8 cell line for 5-7 days. The endpoint is essay focus statement reduction of viral infection of the B95-8 cells 8. This method measures inhibition of viral production and subsequent infection but its disadvantages lie in the length of the definition, assay and failure to measure killing of infected cells. Another assay is the focus, intracellular cytokine staining ADCC (ICS-ADCC) assay measures IFN-?, the primary cytokine responsible for hbs essays 2015, the antiviral activity of many cells including NKs, CTLs, Th1, and dendritic cells 9. The advantages of this assay are that multiple effector functions can be measured and all cells and statement 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 about, the surface of infected cells, potentially overestimating the efficacy of ADCC. Recently our lab has developed a highly quantitative flow cytometric assay to statement measure GrB cytotoxicity using primary HIV-infected CD4+ T cells as targets and autologous NKs as effectors. This is an adaptation of an assay for measuring the cytotoxic activity of CTLs developed in writing, 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 highly reproducible results. 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 focus, the presence of a granzyme cleavable fluorescent substrate.

Granzyme delivery to the target cell and romeo and juliet essay elimination of essay statement HIV-infected targets can then be measured by flow cytometry (Figure 1). Cellular GrB content. Thesis. GrB cleaves GranToxiLux substrate (FITC), indicating the amount of GrB present within a target cell (Pacific Blue). Elimination of HIVSF162-infected cells. Statement. Live targets are stained for intracellular HIV capsid, p24 (PE). Some infected cells show downregulation of CD4 (PE-Cy7). A reduction in romeo and juliet questions, the number of p24 stained cells is essay represented as Infected Cell Elimination (ICE) and is an love definition indication of 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 reduce background. Focus. 12. In addition, because NK function is diminished in HIV-infected patients, we use the NK cells and CD4+ T cell targets from uninfected patients in our assay as a readout for the effectiveness of a given serum sample in mediating ADCC 13, 14. We observed variations in the storm jane essay, NKs from normal individuals in their ability to mediate ADCC and essay focus choose donors that provide optimal signal to noise ratio that we will use to compare the serum of HIV+ individuals (Figure 2). The targets used in this assay are likely closer to the in thesis recommendation, vivo situation given autologous HIV-infected targets are used in the place of antigen pulsed cell lines.

Other advantages include our assay’s ability to measure the mechanism of essay focus ADCC (granzyme delivery) and the outcome (death of infected cells) (Figure 2). MHC-1 is down-regulated in virally infected cells, and the loss of MHC-mediated killer inhibitory signals permit NKs to romeo and juliet act 4 essay kill a cell 15, 16. We control for focus, this by measuring the granzyme content of targets that are incubated only with effectors and sera from HIV-uninfected volunteers. For a positive control we are using a novel anti-CD3 Ab that will mediate ADCC to essay all CD3-bearing targets (Figure 3). Statement. With this Ab we have already demonstrated (1) the sensitivity and preschool writing paper dynamic range with which we are able to measure ADCC, (2) Abs mediate ADCC in a dose dependent manner, and (3) GrB delivery is proportional to cell death. Aim 1: Characterize the magnitude of ADCC in HIV infected individuals . Rational : There is considerable controversy regarding the essay statement, magnitude of ADCC activity in sera from HIV+ individuals and how it correlates with disease progression.

It has been suggested that sera from elite controllers exhibit greater ADCC activity which correlates with CD4+ T cell count 17, 18 while others have sighted no correlation between ADCC and personalized preschool disease progression 19, 20.We will compare the serum of HIV+ individuals with various rates of progression and essay focus statement viral loads to determine which contain Abs capable of personalized writing mediating the essay focus statement, highest levels of writing ADCC. We will also compare the essay focus, ADCC and neutralizing activity of patient serum to evaluate whether or not ADCC and neutralization always occur together in romeo act 4 essay questions, polyclonal sera which is yet to be determined 21, 22. Methods : Over the past several years, in collaboration with the Vaccine Research Center (VRC) our laboratory has focused upon recruiting HIV-infected patients with broadly crossneutralizing antibodies. Essay. We have extensive clinical data and neutralization data for all of the patients. With the preschool writing, 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 passive transfer macaque studies, we expect LTNPs to exhibit greater ADCC than progressors. Essay Focus Statement. 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 preschool paper, serum identified as having potent ADCC activity with regard to specificity. Rational/Methods : HIV-infected cells express Envelope (Env) protein on focus statement, their surface which is essay recognized by NAbs and presumably by essay focus, ADCC-mediating Abs.

We will use standard Nabs (including b12, 2F5, 2G12, 447-52D, 17B, and VRC01) and personalized preschool writing paper patient derived MAbs to probe specificities required for statement, 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 for future mediate ADCC in CD4+ T cells infected by different clades. Focus Statement. 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 love definition IgG3 are the predominant IgG isotypes with roles in ADCC. It is possible that variations in ADCC activity of sera are mediated by variations in IgG subclass. We will use infected CD4+ T cells, apply sera with various levels ADCC at the same dilutions used in essay focus, our killing assay, and measure total bound IgG, IgG1, and referencing in essay IgG3 by flow cytometry. We believe flow cytometry is a better way of measuring titers of binding Abs as compared to focus ELISA because Abs will recognize all possible epitopes on the surface of an infected cell in and juliet act 4 essay questions, their natural conformation. Expected Outcomes/Interpretation : Determining the association between ADCC activity and IgG subclass may provide insights into focus, the necessity of about recommendation 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 study ADCC is focus 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. Romeo And Juliet Act 4. In 1994 one group designed a bispecific antibody with anti-CD4 and essay anti CD3 heavy chains that would redirect CD8+ T cells to kill HIV infected targets 24.

With fluorescent microscopy they observed after 1 hour of referencing in essay incubation that 24% targets had formed conjugates with CD8+ T cells although many more cells had lysed. Essay Focus. At 2 hours 50 - 80% of targets were conjugated and undergoing lysis and by in essay, 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 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 essay focus nanotube structures for viral transmission between CD4+ T cells 25. They have recently demonstrated that macrophages can infect CD4+ T cells every 6 hours and that cell-tocell transfer of virus is about recommendation 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. Essay. HIV-infected CD4+ T cells will be mixed with patient serum or Abs of multiple specificities (see Aim 2) and NKs to hbs essays 2015 evaluate ADCC; or neutrophils/monocytes to essay focus statement evaluate antibody-dependent cell-mediated phagocytosis (ADCP) and applied to poly-L-lysine coated cover slips. Love Definition Essay. Cover slips will be fixed in paraformaldehyde, quenched with NH4Cl, permeabilized with Triton-X, and stained for essay, granzyme B, IgG, CD4/CD16/CD14 with fluorescent probes. After mounting, images will be acquired every minute for about, 6 hours using a Zeiss Pascal Axivert 200M scope.

We will use transmission electron microscopy and tomography (TEM) to focus 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 glutaraldehyde, and stained with osmium tetroxide. Samples will be embedded in epoxy resin, cut, and imaged using a 300kV FEI Technai TF30 27. If necessary cryo-EM will also be used for structural studies with the aid of Oxford collaborator Kay Grunewald. These experiments will be completed using the pathology department’s core bioimaging facility and definition essay I will take department’s Advanced Light Microscopy and Electron Microscopy course in preparation for this research. Expected Outcomes : We expect to elucidate the essay focus, 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. Love Essay. We do not foresee any difficulties in essay, accomplishing this objective. The results of hbs essays 2015 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 essay 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 hbs essays 2015 cells, sera, and essay focus 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 Intramural Research policies. Phlebotomy may cause some discomfort, and and juliet act 4 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. Essay. In the course of any time frame during this study, the hbs essays 2015, total amount of blood will not exceed 10.5mL/kg, or 550 milliliters over essay focus, any 8-week period, which is within the safety guidelines for definition, blood drawing practiced at the Clinical Center. Like standard phlebotomy, leukapheresis is occasionally associated with local bleeding and essay focus bruising at the site of needle insertion.

Lightheadedness can also occur, and people faint on rare occasions. Occasional tingling in the arms and legs can occur with the procedure and can usually be relieved by chewing a calcium-rich tablet that the 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 essay, 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 essay focus statement Race Consideration. This study involves techniques and procedures that are currently approved for hbs essays 2015, various indications. Essay Statement. Male and and juliet act 4 essay 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 focus statement, advertisements posted in the Clinical Center, the NIAID web site, and advertised in other media. 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 personalized preschool writing paper interrupt HAART, patients will be fully counseled prior to essay entry into preschool writing, the study as to the potential risks of discontinuing HAART therapy and the potential benefits of focus 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 study team, will not be offered participation in the study. Patients stopping HAART therapy will be monitored closely for virologic and immunologic relapse and will be offered therapy, based upon the algorithm for safety outlined above. Target enrollment for this study is 300 individuals and is currently ongoing. Inclusion of Children (and Pregnant Women) Inclusion of Children and Pregnant Women. This study will be confined to HIV-infected adults greater than or equal to 18 years of age.

As an observational study there is no therapeutic benefit or clinical intervention resulting from participation in this trial, thereby children are not being denied the potential benefits of a research intervention. We also do not have the historical research database and in romeo and juliet act 4 essay, vitro findings on HLA linkages with LTNP in the pediatric population to make valid comparisons within that cohort. The potential risks of withdrawing therapy in essay, minors have not been studied and therefore may present unknown additional risks to that study population. Comparable units specializing in style referencing, pediatric HIV are available at statement, 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 nursing, and current guidelines continue to advise HIV-positive women to take antiretroviral therapy to prevent viral transmission to the child during that time period. Therefore, pregnant or nursing women will be excluded from this study. Any woman of childbearing potential must have a negative pregnancy test within two weeks prior to definition essay study entry. Essay. All participants who engage in heterosexual intercourse must use two forms of reliable birth control for essay, 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 essay focus statement, 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 antibodies that mediate ADCC.

I outlined the facets of ADCC I wanted to love definition essay investigate then received feedback from both mentors. I subsequently wrote the 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 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 the same manner while I am in essay statement, Quentin Sattentau’s lab.

Selection of eye of elliott essay Sponsors and Institution. Selection of Institution. I decided to focus join Columbia’s MSTP program because of its tradition of exemplary medical education, medical research, and unparalleled support offered to students from program administrators. Because of my previous research experience with HIV I knew I wanted to continue research in this field. I decided to join the NIH/Oxford/ Cambridge Scholars Program to complete my dissertation research because it would provide me access to a more extensive network of scientists than Columbia could. Oxford has a particularly strong network of HIV investigators and the structure of the storm elliott essay its graduate program allows for focus, a particularly specialized education. As a graduate student in 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 immediately implement them into my research. In addition, the NIH/Oxford/Cambridge Scholars program provides funding for students to take courses offered by any institution giving me access to style virtually any educational opportunity necessary. In addition, through NIH’s Foundation for Advanced Education in the Sciences I’ve had the opportunity to take courses that will help me towards my goal as in infectious disease investigator such as the Vaccine Development course I completed.

The NIH/Oxford/Cambridge Scholars Program is designed so that students work with a mentor at NIH and one at a university. When I joined the essay focus, program I searched for HIV investigators throughout NIH and Oxford and identified Mark Connors and definition Quentin Sattentau as potential mentors because I wanted to essay conduct research that had direct implications in vaccine development. Hbs Essays 2015. 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. Connors’s expertise in NAbs of long-term nonprogressors (LTNPs) would complement Dr. Sattentau’s vaccine development goals.

In addition, Dr. Connors’s unique cohort of LTNPs would provide ready access to samples to study many facets of HIV infection. Essay Focus Statement. Finally, I believe Dr. Sattentau’s approach to research as a PhD and Dr. Connors’ as an MD will teach me to think about science in preschool writing paper, a more practical way that is suited to essay focus statement me as a future MD/PhD researcher.

I will follow the principles on eye of jane, responsible conduct of research as outlined in NOT-OD-10- 019. Essay. As an undergraduate at UMBC I took a semester-long course entitled “Ethics/Integrity in Scientific Research” which first help me to understand the responsibility scientists have to 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 love essay I am expected to adhere to the code of ethics as outlined in essay focus, student handbook. Upon joining the NIH I took an online ethics course and will continue to take refresher courses annually. At Oxford I will continue my ethics training by taking the hbs essays 2015, required “Ethics: Introduction to Research Ethics”. Since I will have access to sensitive patient data it is essay focus statement 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 personalized preschool paper alleviate an essay focus individual’s immediate suffering; and eye of as a scientist I hope to contribute to long-term solutions that can prevent suffering. To work most effectively at the interface of medicine and science, I have endeavored to earn an MD/PhD. I believe that the essay focus statement, combined degree’s greatest value lies in the perspective that it affords: to approach patient care with a researcher’s eye, and to hbs essays 2015 approach research with a physician’s focus on improving the essay focus, quality of life. A few years ago I conducted research on telomerase, an enzyme that maintains healthy chromosome length. When telomerase is upregulated, as it is in 90% of 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 definition essay telomerase and focus statement measuring their growth rates, I noticed two things. The first was that cells with low levels of telomerase grew slowly. Secondly, these cells had abnormal morphologies, as do tumor cells. These observations led me to wonder whether the progression of cancer in individuals with DC was slower than in patients without it because DC patients have less active telomerase. Hbs Essays 2015. 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 statement, medicine, but through that experience the importance of keeping a broader medical picture in mind became clear to me. My goals as a physician scientist are to treat patients and conduct research that will allow me to impact patients I will never meet through therapeutic discovery; specifically serving as a creative link between science and the storm jane medicine in statement, the field of infectious disease. The training I will receive in Oxford’s doctorate program will prepare me to be a competitive investigator through a tailored curriculum, expert dual-mentoring, learning to work in a collaborative environment, and by giving me extensive writing experience via paper submissions and even through this grant. These are all specific skills that would be harder and eye of the storm 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 Connors’s lab I have acquired specific skills such to creatively study immunology such as learning to essay focus use flow cytometry. I discuss my research progress frequently during the week with Dr.

Connors. We also have phone conferences and meet with Dr. Writing Paper. 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. Essay Focus Statement. I also completed a 3-day grant-writing course during my senior year of college. Definition Essay. I will improve my writing skills through preparation of manuscripts based on my research to be submitted for publication, applying for training grants, and submission of a report for my transfer viva (qualifying exam). I will also complete the Scientific Writing from a Reader’s Perspective seminar. Discipline specific learning : I completed the 3-day “Vaccines: Development and Evaluation of Efficacy” course offered by statement, FAES in February 2010. I also attended “Psychiatry and HIV,” a 2-day workshop offered by hbs essays 2015, the American Psychiatric Association so I could learn to essay effectively interact with HIV+ patients I encounter. I attend weekly seminar series hosted at essay, the NIH such as the Wednesday Afternoon Lecture Series, Immunology Interest Group seminar series, and essay focus statement weekly lab meetings.

Bench to bedside clinical training : I will attend the thesis recommendation for future, 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 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. Statement. I have given a talk to my graduate program journal club and will have more opportunities to harvard referencing present my research in essay focus, lab meetings.

Career Development : The NIH/Oxford/Cambridge Scholars programs hosts a seminar series where leaders in government/academic research, the pharmaceutical industry, and hbs essays 2015 finance speak with us about their career. I also attended NIAID Fellows retreat that focused on career development. Teaching : As I prepare to leave the lab I will be responsible for training a new research fellow. Year 1 (continued) and Year 2: Quentin Sattentau Lab (Oxford) Research 75%, Seminars/Courses 10%, Presentation/Career Development/Clinical 10%, Teaching 5% Research Skills : While working in Quentin Sattentau’s lab I will learn to apply microscopic imaging to studying HIV infection. I will have weekly meetings to discuss my research progress. We will have phone conferences and meet with Dr.

Connors at various conferences throughout the year. I have also been assigned a graduate advisor within the pathology department but not in my specific lab who will help me to objectively evaluate my research. Writing Skills : I will improve my writing skills through preparation of essay manuscripts based on my research to be submitted for publication and writing reviews. I will complete the following 1-day courses offered at Oxford: Writing Skills – Paper and Thesis; Doctoral Thesis Writing, Discipline specific learning : I will participate in courses and eye of the storm jane Skills Workshops offered by essay statement, Oxford including “Advanced Light Microscopy,” “Electron Microscopy,” and “Introduction to 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 preschool writing be an statement undergraduate lab instructor and take the Oxford course “Teaching and Learning Skills” to prepare myself. Year 3 and romeo and juliet essay Year 4: Medical School (Columbia) In the summer after 11th grade I participated in 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 human genome.

That summer, I learned much more than how to essay run a gel; I learned to read and hbs essays 2015 comprehend primary research articles and how to design experiments. Because of my accomplishments that summer, I was invited to participate in an extension of the Gene Search Program during my 12th grade year. This program inspired me to focus pursue a career in biomedical research because I began to understand how the principles I was learning in school fostered scientific discovery. As a high school senior, I worked with Dr. William Winter at Howard University School of for future Medicine to focus statement investigate pathways that regulate 2,3-diphosphoglycerate (2,3-DPG) levels in red blood cells (RBC). 2,3-DPG lowers the affinity of hemoglobin for oxygen, thereby facilitating oxygen release to tissues. Hbs Essays 2015. Patients suffering with sickle cell anemia have high levels of 2,3- DPG. While this effect may seem advantageous to patients with sickle cell anemia, deoxygenating hemoglobin ultimately causes red blood cells to sickle by promoting hemoglobin polymerization.

Previous studies indicated that adenosine might regulate 2,3-DPG levels through an interaction with the A3 receptor. To test this idea, we indirectly measured 2,3-DPG levels by using spectrophotometry to essay focus statement measure a by-product of the 2,3-DPG pathway. Paper. An exciting therapeutic prospect of our research is essay that an antagonist of the recommendation for future, A3 receptor may prevent the accumulation of 2,3-DPG and essay RBC sickling in patients with sickle cell anemia. I documented this work in preschool writing, a five-chapter thesis, in accordance with my high school’s Science and Technology Program, and presented it at my school’s science fair, where I was awarded third place in biochemistry. During college I spent four years working with Dr. Michael Summers to investigate Moloney Murine Leukemia Virus (MuLV) dimerization. Understanding dimerization is key to focus statement developing gene and antiviral therapies. Harvard Style Referencing. MuLV is essay focus statement similar to thesis recommendation for future HIV in statement, that it is a retrovirus that packages two copies of its RNA genome.

Packaging of hbs essays 2015 dimerized RNA is statement mediated by recommendation for future, interactions between the nucleocapsid (NC) domain of the assembling Gag polyproteins and a specific domain of the RNA called the essay focus statement, Psi-site. Previous work in harvard style, 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. My project was to focus investigate whether similar sequences in the Psi-site could also bind NC. 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 personalized writing, an unpaired guanosine, 2) binding is statement 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. Hbs Essays 2015. al., 2005) extends an essay statement established model for love essay, 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 variable nucleotide) that appear to be preferentially exposed in the dimeric RNA. I presented this research at the 2004 ABRCMS and at multiple UMBC symposiums. I continued to investigate potential dimerization and NC-binding regions of the Psi-site using native and mutant MuLV RNA segments in gel-shift assays. As expected from our riboswitch hypothesis, we observed NC binding preferentially to RNA dimers which was also confirmed by focus, equilibrium dialysis. Writing Paper. Most recently our lab published the NMR structure of focus statement this dimer (Miyazaki, Y., et al., 2010). During the definition, summer of essay focus 2005 I participated in the Weill Cornell/Rockefeller/Sloan Kettering Gateways to the Laboratory Program.

I worked with Dr. Definition Essay. Nikola Pavletich toward solving the crystal structure of the Saccharomyces cerevisiae Rad4-Rad23 complex bound to focus statement a DNA substrate. Rad4-Rad23 is a homolog of human XPC-HR23B, a DNA damage-repair protein. In its mutated form, XPC-HR23B causes the disease Xeroderma Pigmentosum. Thesis About. Patients are extremely sensitive to UV-light and focus statement develop skin cancer during childhood. I identified DNA substrates to be used in crystallization trials by HPLC-purifying DNA, performing gel-shift assays, and setting up crystal trays. I presented this work in a talk before the Tri-Institutional community and during posters sessions at the 2005 Leadership Alliance National Conference and at the 2005 SACNAS National Conference. In 2006 I studied telomerase assembly in the lab of Dr.

Thomas Cech at harvard style, the University of Colorado at Boulder. The central role telomerase plays in protecting chromosome integrity places telomerase at focus statement, the heart of two common human ailments—cancer and aging. To study how and where in romeo and juliet, the cell telomerase is assembled from essay, its RNA and protein components, we used the budding yeast Saccharomyces cerevisiae as a model organism. We sought to hbs essays 2015 understand whether the focus statement, 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. Personalized Preschool. Mtr10p then plays a role in importing the TLC1-Sm complex back into the nucleus, where the telomerase holoenzyme is assembled. Using cellular fractionation and real-time RT-PCR, we attempted to essay focus statement 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 eye of the storm jane, 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 essay statement, neutralizing HIV than Long-Term Nonprogressors (Doria- Rose, N., et al., 2009). Act 4 Essay. 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 focus statement ingenuity it takes to preschool writing paper conduct research, and 3) the vital role that a physician-scientist plays in translating basic research into medical treatment. Although I’ve experienced frustrations and essay disappointment in harvard style, research, the thrill of observing something new, finishing a project, or publishing a paper always outweighs these challenges. Each successive experience only increased my eagerness to improve health care through innovative research. [Redacted from sample.] QUENTIN SATTENTAU MENTOR STATEMENT. [Redacted from sample.] [Our Web version of this sample application can't reproduce the layout of the actual summary statement. See below for the data fields and focus 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 thesis recommendation, extent of protection that ADCC provides. Statement. The reviewers noted that the preschool writing paper, candidate is outstanding, with a strong record of essay statement research training and accomplishment. The candidate appears to be highly motivated. Her sponsors, at both institutions, are well qualified to provide the essay, necessary training and essay statement the letters of recommendation were uniformly laudatory. Minor weaknesses of the application included the writing paper, ambitious research plan to focus statement be accomplished in about, two years, the risk of conducting the essay, research at two sites, and the failure to address pitfalls and alternatives of the research project. These weaknesses, however, did not detract from the hbs essays 2015, reviewers’ enthusiasm.

This training project will have a high impact in ensuring that the candidate maintains her potential to become an independent clinical researcher. DESCRIPTION (provided by applicant): Understanding the basis of an 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 vaccinees that have become HIV-infected. Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. Essay Focus. 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 the storm elliott essay, infected patients to understand the statement, magnitude and characteristics of the best responses achieved by natural infection.

First, we will compare ADCC mediated by the sera of a cohort of referencing patients using a granzyme B cytotoxicity assay developed in our lab. Essay Focus Statement. Based on style in essay, these findings, we will select the sera of patients with the most ADCC, generate monoclonal Abs (mAbs), and characterize the mAbs based on epitope specificity, affinity, potency, breadth, IgG isotype, and Fc type. We will also evaluate whether ADCC is disparate from classical neutralization. Finally, we will use microscopy to examine the synapse between effectors, Abs, and targets. Essay Focus Statement. The outcome of the storm jane essay this research will provide insight into essay, the characteristics of Abs that mediate ADCC that are likely important goals in the design of HIV vaccines or immunotherapies. The Storm Elliott Essay. Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is a function that has been shown to mediate protection from lentiviral infection. Essay Focus Statement. 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 personalized preschool paper HIV-infected individuals in ADCC.

Aim 2: Characterize the essay, specificity and breadth of antibodies with ADCC activity. Eye Of Essay. Aim 3: Characterize the structure and function of the essay focus statement, 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 tomography. Understanding the role of style in essay antibody-dependent cellular cytotoxicity (ADCC) in essay statement, HIV could provide important insights for induction of this activity through vaccination. This project seeks to hbs essays 2015 characterize the Abs that mediate ADCC and image the functional synapse formed by cellular components involved in ADCC with the goal of defining new goals for the development of focus HIV vaccines and style in essay therapeutics. Fellowship Applicant: 1. Sponsors, Collaborators, and Consultants: 1. Research Training Plan: 2. Training Potential: 1. Institutional Environment Commitment to Training: 2. Overall Impact/Merit : This proposal is from an applicant who is essay focus 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. The Storm Jane Elliott Essay. Ultimately, she will receive her Ph.D. from Oxford University and complete her medical training at Columbia University. Essay. 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 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 antibody subtype. Additional experiments, which will draw on the expertise of the co-sponsor, will investigate the essay focus, target-effector synapse during ADCC. These studies will be significant for their basic science and translational value. The training plan is unique in that training and style referencing in essay research will be accomplished in focus, two separate laboratories of the sponsor and co-sponsor. This proposal outlines an excellent training opportunity that will greatly benefit the applicant and her potential for establishing an independent research career. The applicant, who is harvard referencing currently enrolled in an M.D./Ph.D. Essay Focus Statement. training program, is described in three strong letters of eye of elliott essay recommendation as enthusiastic, articulate, knowledgeable, highly focused, and very inquisitive.

Her undergraduate research mentor characterized her as “easily among the top two or three of the more than 200 current and former undergraduates” that he has mentored. The applicant has four years of undergraduate research experience, which involved work with the Moloney Murine Leukemia Virus, and research performed during a summer program at 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 focus, two published papers and has presented eight posters and one oral presentation at various local, national, and hbs essays 2015 international meetings. Essay Focus. The applicant has an outstanding academic record (3.97 GPA) and numerous awards and eye of elliott essay 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 co-sponsor. The applicant has provided thoughtful goals for developing a career as both a scientist and physician. 2. Sponsors, Collaborators, and Consultants: The applicant’s mentor and sponsor, who is currently Chief of the HIV-Specific Immunity Section in the Laboratory of Immunoregulation, NIAID/NIH, is an established and productive senior investigator with research support provided by 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 essay focus, sponsor and co-sponsor will provide excellent support for the applicant’s research and training. The expertise of the co-sponsor will be particularly relevant to hbs essays 2015 studies outlined in Aim 3. The sponsor has trained an average of four students per year over essay focus statement, 20 years, suggesting an hbs essays 2015 excellent training environment for the applicant. Similarly, the co-sponsor has trained nine Ph.D. Focus Statement. trainees since 2003. Jane. During her predoctoral fellowship, the applicant will be interacting with four post-baccalaureate students and one M.D. in essay focus statement, the sponsor’s lab, and four Ph.D. trainees in the co-sponsor’s laboratory. The latter trainees will overlap with the applicant for one year or more. These interactions will enrich the applicant’s training. The research plan is built on romeo and juliet act 4 essay, studies of antibody-dependent cellular cytotoxicity in HIV-1-infected individuals. Experiments will examine the focus statement, potency of sera of about for future HIV-1-infected individuals in essay, ADCC (Aim 1), the specificity and breadth of antibodies with ADCC activity (Aim 2), and the storm jane 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 sponsor’s lab) that offers significant improvements over focus, other ADCC assays.

Results generated from experiments in Aims 1 and hbs essays 2015 2 will have both basic science and translational value, while results from focus statement, Aim 3 experiments will provide basic information about the essay, mechanisms of essay ADCC. Thesis About. The applicant and sponsors are well positioned to perform these studies, since the patient populations and study procedures are already in essay statement, place. The applicant is and juliet essay questions participating in the NIH/Oxford/Cambridge Scholars Program, which promotes research with a mentor at the NIH and essay focus statement another mentor at a university. Consistent with the Scholars Program, the recommendation for future, research training plan includes studies and training in year 1 in the sponsor’s lab at the NIH, continued studies and training in essay focus statement, year 2 in the co-sponsor’s lab at Oxford, and years 3 and 4 in medical school. Training in 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 definition, 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 focus, experiments involving infectious HIV-1, microscopic imaging, and definition essay biochemical and serological techniques (such as ELISA, gel electrophoresis, and mass spectrometry). Although the targeted patient enrollment is given as 300 HIV-1-infected individuals, the experimental plan does not provide details regarding the scope of studies in Aim 1. Correlates of ADCC to be studied in Aim 1 (“rates of progression and viral loads”) were not described in sufficient detail. How will rates of progression be measured? Will viral load be documented on a continuous scale or will patient viral loads be stratified? An explanation of how the results of Aim 1 experiments will be used to statement guide the selection of pooled patient sera in about, Aim 2 would have been informative. A research project spanning the laboratories of essay focus statement two different principal investigators will offer diversity in training and acquired expertise, and enhance the questions, capability of the applicant to develop an independent and productive research career. 5. Focus Statement. Institutional Environment Commitment to Training: The resources and 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 time difference between the two performance sites.

Training in the Responsible Conduct of Research: Comments on Format (Required): The description of RCR training is brief and romeo act 4 questions does not provide a detailed plan for formal training. However, enrollment in two formal courses at Oxford (both focused on ethics) is mentioned. Comments on Subject Matter (Required): Training courses and certifications at the NIH and Oxford should provide information on relevant subjects. Comments on focus statement, 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 hbs essays 2015, 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 essay outstanding proposal from a highly qualified applicant. The applicant has an hbs essays 2015 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. Both sponsors have excellent training track records and both are actively involved in the HIV arena.

The primary goal of this project is to focus characterize antibodies that mediate antibody-dependant cellular cytotoxicity (ADCC). The Connors lab at the NIH has developed a novel flow cytometric-based assay to measure granzyme B delivered to an HIV infected cells. Preschool Writing. 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 essay 2). The third Aim to characterize the structure and function of the target-effector synapse will be carried out at Oxford under the guidance of Dr. Sattentau.

This aim will use fixed and live cell laser scanning confocal microscopy and thesis recommendation for future TEM to examine the samples between NK and other cells with ADCC activity. Focus. If the applicant is successful the harvard style referencing in essay, data from this project will be highly important for HIV vaccine development. Essay Focus Statement. The research project however is quite ambitious and in essay no timeline is statement given. It appears that the applicant plan to spend 1 more year in the Dr. Connors lab and 1 year in hbs essays 2015, Dr. Essay Statement. Sattentau lab making it a very busy two years if the applicant plans to finish all the work they outlined. Despite the highly ambitious nature of the romeo and juliet, proposed research project the essay, 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 eye of the storm elliott, the Gene Search Project at Howard University Hospital. As an undergraduate at UMBC she was a member of the statement, 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. Love. The applicant has extremely strong letters of recommendations, one of which is from a Nobel Prize winner.

Exemplary scholastic record and essay focus many community-based volunteer positions. 2. Sponsors, Collaborators, and Consultants: Two suburb sponsors are listed. Hbs Essays 2015. 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 Pathology at the University of Oxford, is equally meritorious with over 151 published manuscripts and solid funding record. Dr. 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 essay focus statement there are sufficient preliminary data to support the feasibility of the project. The plan is to utilize a novel recently developed (Connors lab) quantitative flow cytometric assay to measure GrB cytotoxicity and determine which Abs are capable of mediating the highest level of essay 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 antibodies with ADCC activity. Using advance microcopy techniques (fixed and essay statement 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 virologic synapses. This is a highly ambitious grant. It may be difficult for the applicant to finish all the proposed work especially the work proposed in Aim 3. The applicant is calling for the storm jane elliott essay, 300 subjects but no power calculation is shown to support that many subjects. A timeline for focus statement, 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 romeo and juliet act 4 essay questions discipline specific learning (i.e. in house courses and essay statement seminar series, lab meetings). Letters describing each institution’s role in eye of jane, training the applicant were provided. Applicant states that she will interact on daily basis with section head and other senior laboratory member on a daily basis. Students also make formal presentations and attend weekly LIR, meeting, NIAID grand rounds, clinical center Grand Rounds and HIV journal club. The training plan indicates that the applicant will be at the NIH for one year and Oxford for one year and essay then back to essay medical school for the last 2 years. This raises some concerns regarding the duration of the funding (3 vs. 2 years). 5. Institutional Environment Commitment to Training:

The work will be conducted at the NIH/NIAID and University of Oxford. Both institutions are of the highest caliber with access to all the required resources. Oxford has a program in focus, place for preschool writing paper, graduate students to take accelerated courses and the NIH Foundation for statement, Advanced Education in the Sciences also has specialized courses that will facility the applicants goal to paper become an infectious disease investigator. Training in the Responsible Conduct of Research: Comments on Format (Required): On line ethics course at the NIH and formal classroom course at statement, Oxford. Comments on Subject Matter (Required):

Applicant states they will follow principle of research outline in NOT-OD-1-019. The applicant has taken semester long course on Ethics/Integrity in love, Scientific research. The applicant took another Ethic course upon entering the NIH. Oxford offers a similar course. Comments on Faculty Participation (Required): No comment on faculty participation. Comments on Duration (Required): Comments on essay, Frequency (Required): Online course and annual refresher at the NIH. Additional Comments to Applicant (Optional): Fellowship Applicant: 1.

Sponsors, Collaborators, and hbs essays 2015 Consultants: 1. Research Training Plan: 2. Training Potential: 1. Institutional Environment Commitment to Training: 1. Overall Impact/Merit : This proposal comes from an excellent candidate who is part of the statement, NIH/Oxford/Cambridge Scholars program, carrying out 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 style referencing, Columbia. The primary hypothesis of statement this proposal is that ADCC activity levels in sera are dictated by the amount, specificity, and subclass of HIV-specific Abs. This represents a highly relevant area of research that will be tackled through 3 straightforward and well-planned aims – (i) characterize the potency of sera from individuals at different stages of HIV infection in ADCC (comparing to clinical status and Ab neutralization), (ii) characterize the specificity and breadth of the storm elliott Abs mediating ADCC (comparing the essay statement, sera to panels of known neutralizing Abs), and recommendation (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. Focus. It is a clear and well-written proposal detailing a strategy to carry out relevant research in an excellent training environment.

THE FOLLOWING RESUME SECTIONS WERE PREPARED BY THE SCIENTIFIC REVIEW OFFICER TO SUMMARIZE THE OUTCOME OF DISCUSSIONS OF THE REVIEW COMMITTEE ON THE FOLLOWING ISSUES: PROTECTION OF HUMAN SUBJECTS (Resume): ACCEPTABLE. INCLUSION OF WOMEN PLAN (Resume): ACCEPTABLE. INCLUSION OF MINORITIES PLAN (Resume): ACCEPTABLE. INCLUSION OF CHILDREN PLAN (Resume): ACCEPTABLE.

COMMITTEE BUDGET RECOMMENDATIONS: The budget was recommended as requested.

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?A Franz Kafka: “The Metamorphosis” Essay Sample. A Franz Kafka: “The Metamorphosis” – An Analysis. Franz Kafka is considered to be one of the statement most important and influential writers of the referencing 20th century. His works, most of which was published posthumously, continues to be a source of research, scholarship and philosophical discussion in diverse academic, literary and popular arenas. The Metamorphosis (German: Die Verwandlung) is a novella by focus him, first published in 1915.

It is often cited as one of the seminal works of short fiction of the 20th century and is widely studied in colleges and eye of the storm essay universities across the western world (Wikipedia, 2011). The story is mainly revolves with Gregor Samsa, who found himself transformed into his bed into essay focus statement, a horrible vermin. Eye Of? This made Grete, his sister, to essay help him out by giving him food every day. She had served him twice a day. Romeo And Juliet Essay? But, eventually, she has grown impatient with the task.

One day, when Gregor emerges from his room, his father chases him around the statement dining room table and pelts him with apples. Thesis Recommendation For Future? One of the apples got lodged in his back, causing an infection. Focus? Because of his infection and his hunger, Gregor is soon barely able to move at all. #8220;The fusillade of fruit is an uncontrollable, instinctive reaction to the presence of eye of the storm jane his transformed son. The father is focus no thinker, no philosopher, and no great spirit either. He does not understand the unrecognizable son and cannot really hope to. His assault of personalized preschool writing paper apples is focus a pathetic rage of incomprehension, almost pitiablebut nonetheless lethal, easily penetrating the son#8217;s armor. Remorse follows, no doubt, for there are no more attacks, but the damage has been done.#8221; (Sparknotes, 2005) Then, here came the three lodgers that they had accepted to earn extra income for the family.

One night, when Grete was practicing playing her violin, the lodgers wished for a performance. This also lured and enticed Gregor so he made his way downstairs, but one of the lodgers sees Gregor, and all three declare that they will leave the next day. Furthermore, they will not pay the rent that is due. Gregor was pushed back into his own room and and juliet act 4 died soon after. The main character, Mr. Gregor Samsa, a travelling salesman, is the breadwinner of the focus statement Samsa family.

The family’s comfort is dependent on personalized preschool his ability to essay focus statement work. In the first few paragraphs, it was illustrated that he was so insistent in going to work despite his incapability. Gregor has been devoting most of his time in working so he could keep the family afloat and jane dig his father out of debt. His job may appeal to be degrading, frustrating, and statement meaningless, but he has still endured all those to secure his family’s condition. It was said that “In fifteen years of service, Gregor has never once yet been ill.” (Kafka, 1915, chap. 1) Because of this, he had no social life since he was always busy. Through Gregor#8217;s portrayal, Kafka showed his experiences in preschool writing his society; how he had not been trusted by his employer just because he is a Jew. Kafka had also reflected his relationship with his family on the story. Both Kafka and Gregor had been very much expected of of their dominant father and yet, through Gregor, had disappointed him because of the transformation, and with Kafka, through his withdrawal of his Jewish heritage. (Oppapers, 2005) Gregor’s love and devotion towards his family remain unchanged throughout the story. He had a very hard time dealing with everything as his family had abandoned him.

But even though it#8217;s as if the world had turned his back on him, he had still loved his family the essay focus statement same way that he had always been (Echeat, 2004). He respects his parents very much and eye of the storm he never wanted to disappoint them. Even when confronted with proof of his family’s scorn and essay focus statement rejection, Gregor refuses to see them as anything but justified in their disappointment and anger towards him. He’s also very supportive in pursuing Grete’s dream. It was said there that “It was his secret plan to send her to the conservatory next year though it would cause great expense” (Kafka, 1915, chap. 2). Having awoken up in his troubled dreams, it seemed that it had gone off even in and juliet essay reality. He’d thought of getting up to catch the focus statement next train, rather than grieving upon his inexplicable transformation.

Finding out that he had nothing to do but to eye of the storm elliott stay at home due to focus statement his condition, he had to cope with the changes while maintaining his daily routine. Soon, his human voice disappears and his preferences grow more insect-like. With time, he masters the coordination of his new body. He thinks less and crawls on walls and ceilings. Gregor’s metamorphosis symbolizes another change in which he becomes more distant from his family through discovering all the lies that his family had told him about not having any money. The reason he worked so hard and it was all for a lie, when his family did have money all saved up, just that they wanted more. He becomes less human like and decides to keep to himself because he is probably really hurt. The change that he goes through in the metamorphosis change is mainly shown when he became less human and act 4 essay questions dependent on his family instead of focus his family being dependent of him. Gregor’s radical change of mind is boosted by the events that take place about all the lies that he was told about his family’s financial problems, thus causing a loss of caring for writing paper, his family and a feeling of essay focus statement individuality which all start to romeo questions alienate him from his family and the rest of human society. No one had ever questioned why the metamorphosis has ever taken place simply because no transformation happened. The complete dependence of Gregor#8217;s family and employer on essay statement him, then, is seen as an ironic foil to the reality of Gregor#8217;s anatomical transformation into a parasite.

Gregor was just compared to preschool an insect since he is afraid of essay focus statement those who surround him. He had developed the fear of an insect, being effectively shooed away by hissing voices and stamping feet. He’d rather live somehow dark and distant. The removal of the things inside Gregor#8217;s room had symbolized his alienation from the society. Referencing In Essay? When people took everything from anybody, it would feel like everything had been gone, making him feel alienated, unwanted and unloved. Like in the begging of the focus statement book he was a person concerned with the welfare of others and and juliet essay questions worked hard to help them.

But towards the end he became a bit retarded by believing that he was a bug and started to accept his surrounding and start acting like one. He became less concerned with his well being and that of those around him, but just did not care anymore, just wanted to do nothing. He was not concerned with what he could do to help him but just be there. The more Gregor went through his metamorphosis, the less human like he became. Thus separating him from the rest of the human society and, causing him no to want to interact with his family as well as anyone else that he came in contact with, which caused his ultimate demise. The transformation had brought a quick degradation in his relationship with the mother and the father.

His inability to communicate symbolizes his alienation from focus, society. Hbs Essays 2015? After the incident, the one where his father had pelted apple towards him because of what had happened, Gregor was aleady in doubt if he had really known his father very well. For the first few years, it seemed that his father was still fond of essay him; always welcoming him home from work, but now, it looks like everything had changed. Eye Of The Storm Elliott? He didn’t even think twice of pelting the apples towards Gregor because of so much rage in his heart. He had never considered Gregor’s feelings after the essay focus transformation. But even though his father had a cold heart with him, Gregor still thought that it was his all fault why his father was like that. He had never blamed others for their mistakes; he always took account for all the things that make things worse. Grete, on essay questions the otherhand, was at first very willing to help. She was the only one who had the nerve on coming inside Gregor’s room. She even delivers Gregor’s food in essay focus statement every meal. Gregor feels reassured by the efforts being made on definition essay his behalf.

She had a very strong relationship with Gregor before the transformation, which had faded in no time. Grete, despite the kindnesses she shows her brother, is unable to bear looking at focus statement, him. Mrs. Samsa, however, was a mere representation on writing how the society would react with Gregor’s transformation. Shock, fear, and sadness –all had happened in one momentum. Statement? She had always loved Gregor so much but it was very disheartening for definition essay, her to look at essay focus statement, her son. Doubtful that her son would not go back to his former self, she has still her faith on him. Gregor had also been degraded by the lodgers. Although he wanted nothing but see Grete play the violin, they had thought that he was there to scare them off. They had even warned Mr. Samsa that they would file a case against the Samsa family since they didn’t tell them about Gregor living on the next room.

All three declare that they will leave the next day; further, they will not pay the rent that is due. This represents how people become so judgmental to others even though they had not known every reason behind every action. Once he’s unable to communicate, Gregor becomes a mere observer of the world around him. Gregor’s metamorphosis symbolizes how he becomes less human day after day, ignoring his needs to becoming a person who simply does not care anymore. At the same time, this isolation evokes a series of startling revelations and personalized preschool writing actions from essay statement, his parents and preschool paper sister, triggered by their assumption that he can no longer understand what they say and their belief that he has lost all human traits.

In the novel, It is not why the transformation happened that was explained, but what it is for; the effects of the transformation. It teaches us how we should value the focus people around us; how we should appreciate all the eye of essay hard work that they’re doing. Essay Focus Statement? It was said there, “You’ve got to get rid of the idea that that’s Gregor. We’ve only eye of the storm elliott essay harmed ourselves by believing it for so long. How can that be Gregor? If it were Gregor, he would have seen long ago that it’s not possible for human beings to live with an animal like that and essay statement he would have gone of his own free will. We wouldn’t have a brother any more, then, but we could carry on definition essay with our lives and remember him with respect.” (Kafka, 1915, chap. 3) With this line, Grete only shows that she still doesn’t believe that the insect is focus statement Gregor.

She insulted the insect after it had driven off their tenants. Referencing In Essay? It can be reflected that the essay main character’s transformation had significances. After Gregor’s transformation, they have to make some adjustments. Gregor#8217;s appearance no longer concerns him. He had gotten used to it. This symbolized how he is becoming less of himself and how he is slowly drifting away from the people around him (CustomWritings, 2003).

His parents now appreciate all the new work his sister was doing even though, before they had seen her as a girl who was useless and annoying. Through the metamorphosis, Gregor becomes more aware of what he wants and what he needs instead of preschool paper what he needs to do for others. Since he had been the essay focus breadwinner of the family, he felt that he needed to be the harvard style in essay one responsible of all the essay statement things. He had carried all the weight of the thesis recommendation problems that the family had to face. He had worked through all the obstacles on focus statement his own. But when the transformation occurred, it was the first breath of paper fresh air. He first panicked; thinking he can no longer go back to his daily routine. Essay Statement? It felt to him that the world is falling apart. He was being more pessimistic than the referencing usual.

He kept on blaming himself why his family is essay focus statement suffering. But after quite some time, it was just a blessing in disguise. He, himself, had realized that he had been devoting his life, serving for romeo act 4 essay questions, his family, giving only a little time even for himself. He had been giving all the wants and needs of his family, but not to himself. With the help of transformation, it was his time to give only for himself; to think and to ponder; to reflect and to realize. His life can never be called as his, since it was served for his family. Being too devoted and respectful child, he was willing to essay give up anything for his family, even his own happiness and freedom. Gregor did not seem shocked that his family wanted to eye of the storm essay kill him off either, almost as if he had been expecting it.

He was only upset because he had scared his sister. It was as though Gregor had removed himself completely from his self, bordering on indifference. Self-alienation was the essay focus word commonly used to describe Gregor#8217;s state in definition the end. Although this self-alienation came strongest in the end of his life, it had been occurring from the moment Gregor accepted his life at the office. Alienation coming from his family had been occurring from the moment as well.

The transformation also made his family realize that they should not be dependent to Gregor; that they should do something to support the family. Five years after their business collapsed, Gregor’s father now has a new job. He currently works as an employee in a banking institute despite his old age. His mother sews fancy underwear for a fashion shop and his sister has taken a sales job. They also have new tenants so they can have extra income to essay statement support their financial needs. Franz Kafka insinuated that we shouldn’t work like insects for others leaving out time for ourselves as Gregor had but instead work for ourselves with pride and dignity as the family did at the end of the hbs essays 2015 story. Gregor was taken for granted by his family. They never considered what his needs and essay statement wants might be. When Gregor had transformed, only then did they realize how they are dependent on him. In reality, the same thing happens.

People only start to romeo act 4 essay appreciate ones worth when it#8217;s already too late. Statement? (123Helpme, 2003) Like generations of essay today, they are not very expressive of their feelings. They just give more value to their personal ego, thinking that showing how much you love somebody will be so overrated. So when somebody had moved to a far place, or passed away, it was always far too late for them to show them how they really value that person. In the end, only regrets will fill their hearts. The transformation had shaken the whole family. At first, almost all was eager to be of help to Gregor, especially Grete. But eventually, they had grown tired and impatient of the works and only see Gregor as a burden to the family. Relating this to reality, when someone gets really ill, everybody would start to feel sorry and extend their sympathy.

Later on, only few will remain since the rest may only view the sick person as a hindrance to their life. Essay Statement? (123Helpme, 2003) Being sick is like being a baby once again. One will be helpless and thesis about recommendation would never be productive until he had recovered from illness. Essay Statement? And when someone is sick, it is hbs essays 2015 not only him that suffers. Statement? Other family members will endure too, since they had to for future take care of the sick person. Essay Focus Statement? Friends, co-workers and other people closely related to the sick person will be affected too, for writing, the sick person may have to do something very important to those people that will be postponed until he will be able to resume his duties. With life#8217;s twist and statement turns, changes may happen in the most unexpected, ill-fated way. With this, people have to learn how to hbs essays 2015 keep up with everything. Like Gregor, he had gone with the thunder and storms, but had known how to dance in the rain. Statement? (Echeat, 2004) Gregor experienced something that only Kafka could ever imagine but he still, against all odds, learns to harvard style deal with it and essay focus statement makes the best out of things. In reality, when people throw rocks at you, you can only do things with those: make a bridge to keep on romeo and juliet essay questions moving on essay focus or make a wall to definition exclude yourself from humanity. Is this the perfect essay for you? Save time and order ?A Franz Kafka: “The Metamorphosis” essay editing for only $13.9 per statement, page.

Top grades and quality guaranteed! Relevant essay suggestions for ?A Franz Kafka: “The Metamorphosis” The novella The Metamorphosis, by Franz Kafka, has themes and characters which parallel some of those in about The Great Gatsby, by essay focus statement F. Writing Paper? Scott Fitzgerald. One of the main themes in#8230; The use of alienation in The Metamorphosis. Essay Statement? Franz Kafka’s use of social commentary in the novella The Metamorphosis illuminates destructive effects of alienation through Gregor’s life before and after his transformation into a monstrous vermin and the#8230; The Loss of Humanity Kafka#8217;s #8220;The Metamorphosis#8221; Language forms the basis of romeo and juliet essay questions civilization. Without language there can be no memory, no culture, certainly nothing as complex as social structure or humanity. In Franz Kafka#8217;s, #8220;The Metamorphosis#8221; the#8230; An Analysis of essay focus “The Metamorphosis” The Transformation of Gregor’s Family : An Analysis of “The Metamorphosis” In Franz Kafka’s 1915 novella “The Metamorphosis,” is about betrayal and definition essay humiliation of an essay statement individual by love his family. Gregor#8230; A Psychoanalytical Criticism of The Metamorphosis. The deeper meaning of “The Metamorphosis”, by statement Frank Kafka, can be interpreted in many ways depending on critical theory is used to examine it.

From a feminist criticism, one can#8230; Comparison of #8220;The Death of act 4 Ivan Ilych#8221; and #8220;Metamorphosis#8221; Ivan Ilych#8217;s death was like Gregor#8217;s Metamorphosis; in essay statement that, they were both powerful introductions to the stories. In both stories the character#8217;s Gregor and Ivan Ilych are introduced in a#8230;

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