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Smalls-Mantey, Sample F31 Application and on family Summary Statement. We are truly indebted to the grantee who allowed us to post this outstanding F31 sample application to help the next generation of investigators write applications. The F31 sample application below doesnt reflect the complex layouts of the actual SF 424 PDF forms. Instead, weve provided a list of key fields from those forms and the applicants responses. In addition, we redacted some applicant-specific information such as contact information, publications in press, and budget specifics. The text of this application is copyrighted. You may use it only for nonprofit educational purposes provided the document remains unchanged and a christmas carol essay the PI, the grantee organization, and functionalist theory NIAID are credited.

Questions? Contact We also offer other outstanding Sample Applications. [Our Web version of this sample application can't reproduce the layout of the actual PDF forms. Here is the summary that eRA Commons generated at the start of the what of the, sample app, which reflects many key fields from that form.] Project/Performance Site Locations. [Our Web version of this sample application can't reproduce the layout of the actual PDF forms.

Performance sites are in theory New York, Maryland, and the United Kingdom.] Research Related Other Project Information. [Our Web version of this sample application can't reproduce the layout of the actual PDF forms. Here are the key fields from that form and the applicant's responses.] Understanding the basis of an immune response that controls infection or provides sterilizing immunity remains a major goal in the search for Mad? Essays 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 functionalist most recent vaccine trials. However, this approach has not been successful in containing viral replication in quality assurance quality vaccinees that have become HIV-infected. Functionalist? 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 quality assurance quality improvement an Fc receptor bearing effector, most commonly natural killer cells (NKs), bind the Ab and use perforin to deliver granzymes which induce apoptosis in the target. We want to study ADCC in infected patients to understand the magnitude and characteristics of the best responses achieved by natural infection. First, we will compare ADCC mediated by the sera of a cohort of patients using a granzyme B cytotoxicity assay developed in our lab. Based on functionalist theory on family these findings, we will select the sera of patients with the most ADCC, generate monoclonal Abs (mAbs), and consultancy report characterize the mAbs based on epitope specificity, affinity, potency, breadth, IgG isotype, and Fc type. We will also evaluate whether ADCC is disparate from functionalist theory, 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 quotes, Abs that mediate ADCC that are likely important goals in the design of HIV vaccines or immunotherapies. Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is a function that has been shown to mediate protection from lentiviral infection. We hypothesize that variations in ADCC activity of sera are dictated by the amount, specificity, and subclass of HIV-specific antibodies. Aim 1 : Characterize the potency of sera of functionalist theory on family, HIV-infected individuals in ADCC.

Aim 2 : Characterize the specificity and breadth of antibodies with ADCC activity. Aim 3 : Characterize the structure and function of the of the, target-effector synapse using both fixed and theory live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM) and tomography. Understanding the Mad?, role of antibody-dependent cellular cytotoxicity (ADCC) in HIV could provide important insights for induction of theory, this activity through vaccination. This project seeks to characterize the Abs that mediate ADCC and image the functional synapse formed by cellular components involved in ADCC with the goal of defining new goals for the development of HIV vaccines and therapeutics. Gallo RC, Salahuddin SZ, Popovic M. Frequent detection and isolation of cytopathic retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS. Mad?? Science.

1984; 224: 500-3. Barre-Sinoussi F, Chermann JC, Rey F, Nugeyre MT, Chamaret S, Gruest J, et al. Isolation of functionalist theory, a T-lymphotropic retrovirus from a patient at a christmas carol risk for acquired immune deficiency syndrome (AIDS). Functionalist? Science. 1983; 220(4599): 868-71.

Organization WH. Expectations Love? 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 theory important in antibody protection against HIV. Nature. 2007; 449(7158): 101-4.

RV144 Phase III HIV Vaccine Trial Press Release. The Crackup Fitzgerald? 2009. Chung A, Rollman E, Johansson S, Kent SJ, Stratov I. The utility of functionalist theory on family, ADCC responses in HIV infection. Current HIV research. 2008; 6(6): 515-9. Kohl S, Starr SE, oleske JM, Shore SL, Ashman RB, Nahmias AJ. Fitzgerald? Human monocytemacrophage- mediated antibody-dependent cytotoxicity to herpes simplex virus-infected cells.

J Immunol. 1977; 118(3): 729-35. Theory? Forthal DN, Landucci G. In vitro reduction of virus infectivity by 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. 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 Mad?, CD8+ T cells is required for 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. Functionalist Theory On Family? Loss of mismatched HLA in a christmas 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 on family, dysfunctional natural killer cell subsets associated with high levels of HIV-1 viremia. Blood. What Cowshed? 2009. Ljunggren HG, Ohlen C, Hoglund P, Yamasaki T, Klein G, Karre K. Afferent and efferent cellular interactions in natural resistance directed against MHC class I deficient tumor grafts. J Immunol.

1988; 140(2): 671-8. De Maria A, Ferraris A, Guastella M, Pilia S, Cantoni C, Polero L, et al. Theory? 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 National Academy of carol essay, Sciences of the functionalist on family, 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 christmas essay a correlation between antibody-dependent cellular cytotoxicity-mediating anti-HIV-1 antibodies and prognostic predictors of functionalist, HIV infection. Is Hamlet? J Clin Immunol. On Family? 2001; 21(3): 227-33. Lambotte O, Ferrari G, Moog C, Yates NL, Liao HX, Parks RJ, et al. Heterogeneous neutralizing antibody and report antibody-dependent cell cytotoxicity responses in HIV-1 elite controllers. Functionalist Theory? AIDS. 2009; 23(8): 897-906. Dalgleish A, Sinclair A, Steel M, Beatson D, Ludlam C, Habeshaw J. 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 quality versus antibody-positive sera. J Virol. 1989; 63(2): 584-90. Doria-Rose NA, Connors M. Antibody-secreting B cells in theory on family 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 was the battle cowshed, void: cell-to-cell spread of human viruses. Nat Rev Microbiol. 2008; 6(11): 815-26.

Chamow SM, Zhang DZ, Tan XY, Mhatre SM, Marsters SA, Peers DH, et al. A humanized, bispecific immunoadhesin-antibody that retargets CD3+ effectors to on family, kill HIV-1- infected cells. J Immunol. What Battle Of The Cowshed? 1994; 153(9): 4268-80. Jolly C, Kashefi K, Hollinshead M, Sattentau QJ. HIV-1 cell to functionalist theory, cell transfer across an Env-induced, actin-dependent synapse.

J Exp Med. 2004; 199(2): 283-93. Groot F, Welsch S, Sattentau QJ. Efficient HIV-1 transmission from macrophages to T cells across transient virological synapses. Consultancy Report? Blood. Functionalist On Family? 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. What Was The Battle? J Virol. 2010; 84(7): 3516-27.

Intellectual Support : My mentors Mark Connors and functionalist Quentin Sattentau are leading experts in their field and quality versus improvement have mentored numerous people that have gone on functionalist on family to successful careers in a christmas carol essay science. This dual mentorship will teach me to aggressively approach science from two distinctive perspectives in my graduate career, something most PhD scientist dont experience until their first post-doctoral experience. While at theory NIH I will have received extensive support of great expectations love, 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 FAES at NIH I have access to theory, courses to improve lab specific skills and foundation knowledge. I have already participated a Vaccine Development course. At Oxford I will take seminars at the Centre for Excellence in Teaching and Learning for report 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 provided me any reagent/equipement need for the proposed research. Lab Space I am provided with a desk and personal desktop computer.

Bench space is shared within the lab. Safety In order to ensure our safety from hazardous blood samples we work under laminar flow hoods in a P2 facility with P3 practices. Samples We receive blood samples from patients enrolled within our protocol. We also have access to functionalist, the NIH blood blank for normal human blood samples. As provided by Quentin Sattentau: Oxford University is held in high international regard for its strong teaching and research background and its unique and personal teaching style. Within the University the quality versus quality improvement, Department of functionalist, Pathology has an excellent reputation for graduate teaching, and consultancy has achieved 100% success in functionalist on family DPhil graduation within 4 years for the past 3 years.

The Department has a strong graduate support structure, in which each student has a principal supervisor, the report, possibility of a cosupervisor, a departmental advisor, and a college advisor. Theory? The supervisor(s) will generally meet with the student multiple times a term, and each of the non-supervisory mentors will meet with the candidate approximately once a term and will file reports on great quotes the candidates progress. The candidate will be expected to attend a series of high profile external seminars that take place once a week in term time, and on family a regular series of internal seminars from Essays, leading and more junior University and functionalist theory on family Departmental investigators and students. Adjoa will attend and participate in the weekly lab journal club, in great expectations which scientific papers are taken apart critically by members of the group, and a weekly laboratory meeting in which each lab member presents their ongoing research. Theory On Family? Adjoa will also be encouraged to chat to other members of the report, department during tea lunch and coffee breaks, a longstanding Oxford institution! The Department has a fully equipped quiet library for literature research and other reading and writing. Adjoa will have her own writing up space close to her work bench, equipped with a portable computer and functionalist theory 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 quality versus quality improvement sorting suite, an electron microscopy facility with full technical and scientific support. We also have strong connections with the biochemistry and chemistry departments and the Weatherall Institute of theory on family, Molecular Medicine at was the cowshed 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 theory associated incubators and a christmas essay 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 on family ancillary equipment, in report which HIV-1 is grown and used to infect cells.

Flow cytometry. The department has a core flow cytometry facility with three BD analyzers (2 x FACScalibur 2 x LSR2) and two sorters. Molecular biology. The Sattentau laboratory has all facilities for functionalist molecular biology carried out under containment levels I, -II and III. Light microscopy. The department has 5 confocal microscopes of varying sophistication and resolution, two of which are set up for assurance versus quality live cell immunofluorescence work. The department has three recently acquired high-speed CCD cameras and associated microscopes for live cell immunofluorescence analysis.

The Sattentau laboratory has a Zeiss Pascal confocal microscope and a new CCD camera set up with a Zeiss Axiovert 200 microscope for fixed and live cell work in the containment-III laboratory, allowing real time analysis of live, infected cells. Electron microscopy. The department has two transmission electron microscopes and a scanning electron microscope, all of which are maintained and run as a central service facility by a skilled dedicated operator. Our collaborator in Oxford (Kay Grunewald) is located at theory on family the University Churchill campus, which is 4 miles from the report, 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. Functionalist Theory? See below for the biographical sketch attachments.] My interest in studying infectious diseases stems from the 2001 anthrax scare in consultancy 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 following summer I came to understand how medical research impacts treatment for routine illnesses and also plays a role in functionalist theory on family 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 Is Hamlet the Gene Search Project and researched biochemical pathways that influence sickle cell anemia with Dr. William Winters at Howard University Hospital. This experience taught me how research and medicine are intertwined and inspired me to become a physicianscientist. At UMBC I was a member of the Meyerhoff Scholars Program and an HHMI Undergraduate Research Scholar where numerous lab experiences, seminars, rigorous classes, and multiple opportunities for scientific presentations prepared me for a career as a scientist.

My lab skills and understanding of a research career increased exponentially during my four years in Michael Summers lab. Not only did I learn about protein purification, gel electrophoresis, NMR, and 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. On Family? Working with Dr. Nikola Pavletich I learned the value of immediately organizing my data so that others could analyze my results quickly. In the lab of Tom Cech I had the responsibility of designing a new protocol to study telomerase assembly, a skill that has enabled me to design a novel assay to investigate AIDS specific questions in my graduate studies. The medical education Ive received at Columbia has given me a unique perspective with which I approach scientific questions. Under the tutelage of Mark Connors and Quentin Sattentau, leaders in consultancy their fields on nonprogressive HIV infection and functionalist on family viral transmission, my understanding of infectious disease and my capacity to pursue further research in consultancy these fields will be greatly enhanced. In addition, the theory on family, courses, medical preceptorships, presentations, teaching, and career development activities I am undertaking in the forthcoming years make me an the crackup fitzgerald 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 theory Outstanding Achievement 2006 Goldwater Scholar 2007 UMBC Top Senior Biochemistry Major 2007 Graduated Sum Cum Laude from UMBC 2007 NIH/Oxford/Cambridge Scholar 2010 International AIDS Vaccine Conference Scholarship 2011 Keystone Symposia Underrepresented Minority Scholarship.

2004 Omicron Delta Kappa Leadership Honor Society 2004 Golden Key International Honor Society, Sophomore/Alumni Director 2007 Phi Beta Kappa. Dey A, York D, Smalls-Mantey A, Summers MF. 2005. Composition and sequence-dependent binding of RNA to the nucleocapsid protein of Moloney murine leukemia virus. Biochemistry. Quality Quality? 44(10), 3735-3744. Miyazaki Y, Irobalieva RN, Tolbert BS, Smalls-Mantey A, Iyalla K, Loeliger K, D'Souza V, Khant H, Schmid MF, Garcia EL, Telesnitsky A, Chiu W, Summers MF. 2010. Structure of a conserved retroviral RNA packaging element by NMR spectroscopy and theory on family cryo-electron tomography.

J Mol Biol. What Was The Cowshed? 404(5), 751-772. Smalls-Mantey A, Winter W. 2003. Biosynthesis of 2,3-diphosphoglycerate in Erythrocytes. Poster presentation at ERHS Science Fair and Prince Georges County Science Fair. Functionalist On Family? March 2003. Smalls-Mantey A, Dey, A, Phillip C, Summers MF. 2004. Characterization of High-Affinity Nucleocapsid Protein Binding Sites Within the Moloney Murine Leukemia Virus RNA Packaging Signal.

Poster presented at 2004 Annual Biomedical Research Conference for Minority Students, November 2004. Expectations Quotes? Smalls-Mantey A, Min J, Pavletich N. 2005. Structural and Biochemical Studies of the Yeast Rad4-Rad23 Complex Bound to DNA. Poster presented at functionalist Tri-Institutional Gateways to the Laboratory Program Symposium and fitzgerald 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 theory on family 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 love 2010 Keystone HIV Vaccines (X5) Meeting. Functionalist Theory On Family? 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. Report? Poster presented at functionalist theory on family 2010 Oxford Pathology Department Graduate Student Symposium. June 2010. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. 2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells.

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

September 2010. Smalls-Mantey A. 2010. Antibody-Dependent Cellular Cytotoxicity and the Control of HIV. Talk given at Frontiers in functionalist on family Biomedicine NIH/Oxford/Cambridge Scholars Program student presentation series. November 2010. [Outside research support redacted from sample.] A. What Was The Of The? Personal Statement Regarding Project. Currently HIV-specific ADCC is considered one of the functionalist theory on family, very top priorities in the field of HIV-specific immunity.

Our laboratory has assembled the best techniques and reagents in the field to carry out this project. We have extensive experience in HIV-specific antibodies and cytotoxicity. In addition we have a long track record of training students to report, prepare them for functionalist theory on family a carreer in biomedical research. 1985 - 1986 Intern in Pediatrics, The Boston Floating Hospital, New England Medical Center, Boston, Massachusetts 1986 - 1988 Resident in Pediatrics, The Boston Floating Hospital, New England Medical Center, Boston, Massachusetts 1988 1989 Chief Resident in Pediatrics, The Boston Floating Hospital, New England Medical Center, Boston, Massachusetts July 1989 - October 1989 Medical Staff Fellow, Respiratory Viruses Section, Laboratory of Infectious Diseases, National Institute of what was the of the cowshed, Allergy and Infectious Diseases, National Institutes of theory, Health, Bethesda, MD October 1989 - June 1993 U.S. Was The Battle? 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 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 Connors M. HLA B*5701 is highly associated with restriction of theory on family, 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 a christmas carol essay, 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 theory, HLA B*5701+ long-term nonprogressors and progressors to restrict HIV replication is not caused by loss of recognition of autologous viral gag sequences. Consultancy Report? J Virol 2003; 77(12); 6889-6898. Iyasere C, Tilton JC, Johnson AJ, Younes S, Yassine-Diab B, Sekaly RP, Kwok WW, Migueles SA, Laborico AC, Shupert WL, Hallahan CW, Davey RT Jr., Dybul M, Vogel S, Metcalf J, and Connors M. Diminished proliferation of HIV-specific CD4+ T Cells is associated with diminished IL-2 production and recovered by exogenous IL-2. J Virol 2003; 77(20); 10900-10909. Functionalist On Family? Tilton JC, Luskin MR, Johnson AJ, Manion M, Hallahan CW, Metcalf JA, McLaughlin M, Davey RT Jr., and Connors M. Changes in paracrine IL-2 requirement, CCR7 expression, frequency, and quality quality 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, OShea MA, Mican J, Packard B, Komoriya A, Palmer S, Wiegand AP, Maldarelli F, Coffin JM, Mellors JW, Hallahan CW, Follman DA and Connors M. Lytic Granule Loading of CD8+ T-Cells is Required for HIV-Infected Cell Elimination Associated with Immune Control. Immunity 2008; 29 (6): 1009-1021. Functionalist Theory On Family? 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 what was the of the cowshed 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 functionalist theory on family sera: clustering analysis and association with clinical variables. Quality Quality? J Virol 2009 84(3): 1631-1636.

Doria-Rose NA and Connors M. Antibody-secreting B cells in HIV infection. Functionalist Theory? Curr Opin HIV AIDS 2009 4:426-430 Wu X, Yang ZY, Li Y, Hogerkorp CM, Schief WR, Seaman MS, Zhou T, Schmidt SD, Wu L, Xu L, Longo NS, McKee K, O'Dell S, Louder MK, Wycuff DL, Feng Y, Nason M, Doria-Rose N, Connors M, Kwong PD, Roederer M, Wyatt RT, Nabel GJ and Mascola JR. Rational design of envelope identifies broadly neutralizing human monoclonal antibodies to HIV-1. Expectations? Science 2010 329:856-861. Migueles SA and Connors M. Long-term nonprogressive disease among untreated HIV-infected individuals: clinical implications of understanding immune control of HIV. JAMA 2010 304:194-201. [Redacted publication in press] The research direction of the HIV-Specific Immunity Section of the Laboratory of on family, Immunoregulation is focused upon mechanisms of Essays, immunity to HIV. The laboratory is staffed by functionalist on family PhD and MD scientists with extensive experience in Is Hamlet Mad? the humoral or cellular immune response to HIV. In addition we have close collaborations with investigators at the NIAID Vaccine Research Center in the area of functionalist on family, 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. 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 the crackup fitzgerald, Office of AIDS Research for work on the humoral response to HIV. Positions and on family Employment.

1985 - 1992 Lecturer, Department of Genito-Urinary Medicine, University College London, UK 1989 - 1991 Visiting scientist, Howard Hughes Medical Institute, Columbia University, New York, NY 1992 - 1998 Staff scientist (Director of Research), Centre dImmunologie, Marseille, France 1999 - 2003 Reader, Section of 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 the Advancement of Science 1998 - Member, American Society for Microbiology 1998 - Member, British Society for Immunology, UK 1998 - Member, Society for General Microbiology, UK 2005 - Member of the Pasteur Institute Paris France Scientific Evaluation Committee. 2005 - Member of the Vaccine Research Center (VRC) NIH Board of Scientific Councilors 2007 - Member of BMGF Vaccine Monitoring Centre SAB 2008 - Member of NIH HIVRAD/Novartis Neutralizing Antibody Vaccine Consortium SAB 2009 - Member of study section for Swedish Program for International Development 2010 - Member of NIH study section for the B cell Immunology Partnership Program for HIV-1 Vaccine Discovery (U19) C. Selected Peer-Reviewed Publications. Most relevant to what of the cowshed, 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 theory sensitive to was the cowshed, entry inhibition. J. Virol. 84: 3516-3527. Theory On Family? Gonzalez N, Bermejo M, Calonge E, Jolly C, Arenzana-Seisdedos F, Pablos JL, Sattentau QJ and was the of the 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 Sattentau QJ. (2008). Efficient HIV-1 transmission from macrophages to T cells across transient virological synapses. Blood 111: 4660-4663. Sowinski S, Jolly CJ, Berninghausen O, Purbhoo, MA, Chauveau A, Kohler K, Oddos S, Eissmann P, Brodsky FM, Hopkins C, Onfelt B, Sattentau QJ and Davis DM (2008). Membrane nanotubes physically connect T cells over long distances presenting a novel route for HIV-1 transmission.

Nature Cell. Biol. 10: 211-219. Jolly C and Sattentau QJ (2007). HIV-1 assembly, budding and cell-cell spread in T cells takes place in tetraspanin-enriched plasma membrane. J. Virol.

81:7873-7884 Jolly C and Sattentau QJ (2007). Adhesion molecule interactions facilitate human immunodeficiency virus type-1-induced virological synapse formation between T cells. Theory? J. Battle Of The? Virol. 81: 13916-13921. Theory? 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 Sattentau QJ. Great Expectations? 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. On Family? J. Exp Med. 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. Is Hamlet Mad?? 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 functionalist theory on family cryo-electron tomography. Consultancy? Plos Pathogens August 2 (8): e83 Sheppard NC, Bates AC and theory on family 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 assurance quality improvement, 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 functionalist on family, original version, these attachments met the page limits.] Understanding the basis of an immune response that controls infection or provides sterilizing immunity remains a major goal in the search for effective vaccines or immunotherapies for HIV.

Antibodies (Abs) induced by candidate vaccines to quality versus, 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. Functionalist On Family? 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 Mad?, mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. In ADCC, Fc-bearing Abs bind viral epitopes coating an infected CD4+ target T cell and an Fc receptor bearing effector, most commonly natural killer cells (NKs), bind the Ab and use perforin to deliver granzymes which induce apoptosis in the target. We want to study ADCC in infected patients to understand the magnitude and characteristics of the functionalist theory, best responses achieved by natural infection.

First, we will compare ADCC mediated by what was the the sera of a cohort of patients using a granzyme B cytotoxicity assay developed in our lab. Based on these findings, we will select the sera of patients with the most ADCC, generate monoclonal Abs (mAbs), and characterize the mAbs based on on family epitope specificity, affinity, potency, breadth, IgG isotype, and Fc type. Consultancy? We will also evaluate whether ADCC is functionalist theory disparate from classical neutralization. Finally we will use microscopy to report, examine the synapse between effectors, Abs, and targets. Theory? The outcome of this research will provide insight into the characteristics of Abs that mediate ADCC that are likely important goals in great expectations quotes the design of HIV vaccines or immunotherapies.

Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is a function that has been shown to mediate protection from lentiviral infection. Theory On Family? We hypothesize that variations in ADCC activity of carol, sera are dictated by the amount, specificity, and functionalist theory on family subclass of HIV-specific antibodies. Aim 1 : Characterize the potency of sera of HIV-infected individuals in ADCC. In ADCC, Abs bind viral epitopes that are presented by infected CD4+ T cells. Consultancy? NKs expressing an Fc receptor bind the Fc domain of the Ab and on family use perforin to deliver granzymes to the HIV-infected cell. A Christmas Carol Essay? Subsequently, granzymes induce apoptosis within the on family, 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 cowshed residual percent of functionalist on family, targets expressing p24, HIV capsid. Compare the serum of HIV+ individuals with various rates of progression and viral loads to determine which contain Abs capable of mediating the highest levels of ADCC. Compare the consultancy, ADCC and neutralizing activity of patient sera. Aim 2 : Characterize the specificity and breadth of antibodies with ADCC activity.

Our laboratory has panels of NAbs derived from patients with known serum neutralizing or ADCC-mediating activity. Determine whether recognition of specific epitopes is required for theory ADCC. Define the breadth of the what battle of the, polyclonal sera by its ability to mediate ADCC in functionalist on family CD4+ T cells infected by different clades of HIV. Titer serum total IgG, IgG1, and IgG3 binding infected CD4+ T cells. Aim 3 : Characterize the structure and function of the target-effector synapse. Using both fixed and consultancy live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM) and tomography, we will examine the functionalist, synapse formed between NK and other cells with potential ADCC activity (macrophages and neutrophils) and infected target cells.

We will specifically investigate: The structure of a functional ADCC synapse. The kinetics of ADCC function in real time and quality versus its relation to functionalist theory, antibody type and specificity. A role for antibody-dependent cell-mediated phagocytosis (ADCP) in the crackup 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 functionalist, AIDS in 1983, HIV infection has reached epidemic proportions with an estimated 2 million people worldwide dying from AIDS each year 1- 3. There is more public awareness about how to prevent HIV infection but with the growing incidence of HIV infection and the limitations of current antiretroviral therapy, there is an imperative need for a vaccine.

One of the most critical areas of research on vaccines for HIV is the search for correlates of immunity. At present the love, only method for determining the potential benefits of functionalist theory on family, a vaccine with regard to assurance versus, protection from infection or lowering of viral load upon infection remains very large clinical efficacy trials. Determining correlates of immunity would tremendously accelerate the development process by permitting some estimation of clinical efficacy prior to a Phase III trial. Reliable correlates would permit screening of vaccine candidates at a much earlier point in development with smaller numbers of theory, participants. Over the past 20 years, research into correlates of immunity to HIV have largely focused on CD8+ cytotoxic T lymphocytes (CTLs) or neutralizing antibodies (NAbs). Mad? Essays? In addition to neutralization, antibodies (Abs) may fight viral infections by functionalist theory antibody-dependent cell-mediated phagocytosis (ADCP) or by antibody-dependent cellular cytotoxicity (ADCC). In ADCC, Abs bind viral epitopes presented on infected target cells and Fc? receptor-bearing effector cells such as natural killer cells (NKs) recognize and bind the carol essay, Fc? region of these Abs. Upon cross-linking of the functionalist theory on family, Fc? receptor, NKs release perforin that punctures the targets cell membrane and a christmas 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 theory providing immunity to quality assurance versus quality improvement, 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. Theory? Prior to consultancy report, 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 on family proof of the concept that ADCC can eliminate lentivirus-infected cells in vivo.

In addition, some recent data in human efficacy trials has also greatly increased interest in ADCC as an important component of vaccine induced immunity to HIV. In September 2009 data from the phase III RV144 (Thai) vaccine trial were released and love quotes showed that the functionalist on family, prime-boost combination of consultancy report, ALVACR HIV and AIDSVAXR B/E reduced HIV infection. Functionalist? This vaccine regimen was known to not elicit a strong cellular or NAb response. It does, however, elicit binding antibodies that have suggested to the field that the protective efficacy is likely mediated through ADCC 5. Report? Our laboratory is part of a large international effort to characterize ADCC antibodies from this trial in order to determine whether there were higher ADCC-mediating antibodies in 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 nature of an functionalist on family effective ADCC response have not been addressed. For example, the number, affinity, and specificities of the crackup fitzgerald, antibodies necessary to mediate ADCC remains poorly understood. Our laboratory has now developed assays that can accurately measure ADCC against HIV-infected cells. Theory? 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 consultancy, provide a more robust assay for HIV-specific ADCC. Past studies of ADCC have been hampered by the availability of assays that are only partially quantitative.

There are four primary methods that people use to measure ADCC 6. On Family? The most prevalent is the consultancy, chromium release assays (CRA), which is highly variable, does not specifically account for the mechanism of killing, and on family is insufficiently quantitative 7. The ADCVI assay incubates targets, serum, and consultancy report effectors for 24 hours and the supernatant is applied to the B95-8 cell line for functionalist on family 5-7 days. The endpoint is what cowshed 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 assay and failure to functionalist, measure killing of infected cells. Another assay is the intracellular cytokine staining ADCC (ICS-ADCC) assay measures IFN-?, the primary cytokine responsible for the antiviral activity of was the of the cowshed, 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 Abs are from the same patient, but it is still a proxy that does not measure HIV-infected cell killing. Functionalist? The rapid and the crackup fluorometric antibody-dependent cellular cytotoxicity (RFADCC) assay directly measures cell killing but requires whole protein 10. Theory On Family? While use of protein pulsed targets is a sensitive method, it likely places artificially high amounts of protein on the surface of infected cells, potentially overestimating the efficacy of ADCC. Recently our lab has developed a highly quantitative flow cytometric assay to measure GrB cytotoxicity using primary HIV-infected CD4+ T cells as targets and autologous NKs as effectors. Versus Quality Improvement? This is an adaptation of an assay for measuring the cytotoxic activity of CTLs developed in on family 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 the presence of a granzyme cleavable fluorescent substrate. Granzyme delivery to the target cell and elimination of the crackup fitzgerald, HIV-infected targets can then be measured by flow cytometry (Figure 1). Cellular GrB content.

GrB cleaves GranToxiLux substrate (FITC), indicating the amount of functionalist on family, GrB present within a target cell (Pacific Blue). Elimination of Is Hamlet Mad? Essays, HIVSF162-infected cells. Live targets are stained for intracellular HIV capsid, p24 (PE). Some infected cells show downregulation of CD4 (PE-Cy7). Functionalist Theory On Family? A reduction in versus improvement the number of p24 stained cells is represented as Infected Cell Elimination (ICE) and functionalist theory is an indication of assurance versus, 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 theory the context of consultancy, MHC mismatch and cause nonspecific killing, autologous cells are used in our assay to reduce background. Functionalist? 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 carol the effectiveness of functionalist on family, a given serum sample in mediating ADCC 13, 14. We observed variations in NKs from normal individuals in their ability to Mad? Essays, mediate ADCC and choose donors that provide optimal signal to noise ratio that we will use to compare the serum of functionalist theory, HIV+ individuals (Figure 2). The targets used in versus quality improvement this assay are likely closer to on family, the in vivo situation given autologous HIV-infected targets are used in the place of antigen pulsed cell lines. Consultancy? Other advantages include our assays ability to measure the mechanism of theory, ADCC (granzyme delivery) and Is Hamlet Mad? Essays the outcome (death of infected cells) (Figure 2). Functionalist? MHC-1 is down-regulated in virally infected cells, and the loss of MHC-mediated killer inhibitory signals permit NKs to what of the, kill a cell 15, 16. We control for this by measuring the functionalist on family, granzyme content of targets that are incubated only with effectors and sera from HIV-uninfected volunteers. For a positive control we are using a novel anti-CD3 Ab that will mediate ADCC to all CD3-bearing targets (Figure 3). Carol Essay? With this Ab we have already demonstrated (1) the sensitivity and dynamic range with which we are able to theory, 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 what was the of the cowshed, ADCC in HIV infected individuals . Rational : There is considerable controversy regarding the magnitude of ADCC activity in sera from HIV+ individuals and how it correlates with disease progression. It has been suggested that sera from elite controllers exhibit greater ADCC activity which correlates with CD4+ T cell count 17, 18 while others have sighted no correlation between ADCC and functionalist on family disease progression 19, 20.We will compare the serum of HIV+ individuals with various rates of progression and viral loads to battle, determine which contain Abs capable of mediating the highest levels of ADCC. We will also compare the ADCC and neutralizing activity of functionalist, patient serum to evaluate whether or not ADCC and neutralization always occur together in polyclonal sera which is report yet to be determined 21, 22. Methods : Over the past several years, in functionalist on family collaboration with the fitzgerald, Vaccine Research Center (VRC) our laboratory has focused upon recruiting HIV-infected patients with broadly crossneutralizing antibodies. Functionalist On Family? We have extensive clinical data and neutralization data for all of the patients. With the data we generate from a christmas carol, our ADCC assay we are able to on family, 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. 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 a christmas carol essay, serum identified as having potent ADCC activity with regard to specificity. Rational/Methods : HIV-infected cells express Envelope (Env) protein on their surface which is recognized by NAbs and presumably by ADCC-mediating Abs. We will use standard Nabs (including b12, 2F5, 2G12, 447-52D, 17B, and VRC01) and patient derived MAbs to probe specificities required for ADCC. An ideal HIV vaccine would be able to protect against diverse virus strains so it is important to theory, determine whether Abs raised against one clade of Mad? Essays, virus would be able to mediate ADCC in CD4+ T cells infected by functionalist on family different clades. We will use multiple strains of virus from clade A/E, B, and C to infect CD+ T cells and compare killing of a christmas carol essay, target cells in theory the presence of serum pooled from Is Hamlet, clade B HIV+ patients. IgG1 and theory on family IgG3 are the predominant IgG isotypes with roles in ADCC. It is possible that variations in fitzgerald ADCC activity of sera are mediated by variations in IgG subclass. We will use infected CD4+ T cells, apply sera with various levels ADCC at the same dilutions used in our killing assay, and measure total bound IgG, IgG1, and IgG3 by flow cytometry.

We believe flow cytometry is a better way of functionalist theory on family, measuring titers of quotes, binding Abs as compared to ELISA because Abs will recognize all possible epitopes on the surface of an infected cell in their natural conformation. Expected Outcomes/Interpretation : Determining the association between ADCC activity and IgG subclass may provide insights into the necessity of eliciting a specific class of IgG via particular antigens and adjuvants. Potential Complications : We now have robust assays for subclass binding and do not expect complications with this portion of the project. Aim 3: Evaluation of the target-effector synapse. Rational : Using microscopy to study ADCC is important because it will help us define structural and kinetic parameters and will relate structure to function.

Multiple mechanisms for cell-to-cell spread of HIV have been described but few have described inhibition of viral transmission or targeted cell death 23. To date there has been no study using microscopy to functionalist, investigate ADCC in HIV using NKs or macrophages as effectors and CD4+ T cells as targets. In 1994 one group designed a bispecific antibody with anti-CD4 and anti CD3 heavy chains that would redirect CD8+ T cells to kill HIV infected targets 24. With fluorescent microscopy they observed after 1 hour of incubation that 24% targets had formed conjugates with CD8+ T cells although many more cells had lysed. At 2 hours 50 - 80% of targets were conjugated and undergoing lysis and by 24 hours all targets had been lysed. However, the bispecific antibody may not be representative of ADCC with naturally occurring HIV-specific antibodies. Assurance Versus Quality? In addition, the functionalist, 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 was the HIV infection. They were the first to demonstrate HIV-1 spread between T cells via virological synapses and nanotube structures for viral transmission between CD4+ T cells 25. They have recently demonstrated that macrophages can infect CD4+ T cells every 6 hours and that cell-tocell transfer of virus is 10-fold more efficient than free virus spread 26. To study the kinetics of ADCC in real time we will use time-lapse laser scanning confocal microscopy (time-lapse LSCM). Functionalist On Family? Cell populations will be negatively selected from PBMCs using magnetic cell sorting. HIV-infected CD4+ T cells will be mixed with patient serum or Abs of Mad? Essays, multiple specificities (see Aim 2) and NKs to evaluate ADCC; or neutrophils/monocytes to evaluate antibody-dependent cell-mediated phagocytosis (ADCP) and applied to poly-L-lysine coated cover slips. Cover slips will be fixed in functionalist on family paraformaldehyde, quenched with NH4Cl, permeabilized with Triton-X, and stained for granzyme B, IgG, CD4/CD16/CD14 with fluorescent probes. Essays? After mounting, images will be acquired every minute for 6 hours using a Zeiss Pascal Axivert 200M scope. We will use transmission electron microscopy and on family tomography (TEM) to generate higher resolution static images of the ADCC functional synapse.

For TEM targets, serum, and effectors will be incubated for approximately 1 hour, fixed in paraformaldehyde and consultancy report 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 departments core bioimaging facility and I will take departments Advanced Light Microscopy and Electron Microscopy course in preparation for this research. Expected Outcomes : We expect to elucidate the kinetic and functionalist theory 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. Great Expectations? We do not foresee any difficulties in accomplishing this objective. The results of these studies will include novel MAb with possible therapeutic value; a new tool for the study of ADCC with potential utility for theory on family HIV vaccine trials; and fitzgerald increased understanding of functionalist theory on family, basic HIV immunology. A Christmas Essay? 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 theory, these important studies because of our large collection of cells, sera, and detailed clinical data from patients that are part of the NIAID HIV clinic, and both labs extensive experience in this area. Samples are collected under the NIAID IRB-approved protocol 02-I-0086. Protections for human subjects are in accordance with NIAID Division of Intramural Research policies.

Phlebotomy may cause some discomfort, and occasionally some bleeding or bruising at the site. On rare occasions, people faint while having blood drawn. Blood drawing will not exceed 150 milliliters (30 teaspoonfuls) on any day or visit. In the course of any time frame during this study, the cowshed, total amount of blood will not exceed 10.5mL/kg, or 550 milliliters over functionalist theory any 8-week period, which is within the safety guidelines for blood drawing practiced at great love quotes the Clinical Center. Like standard phlebotomy, leukapheresis is occasionally associated with local bleeding and theory on family bruising at the site of a christmas carol, 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 theory on family very rare occasions (less than 1 in every 1,000 times), malfunction of the a christmas essay, leukapheresis machine can result in blood loss of theory, 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 fitzgerald, Women and Minorities. Gender, Ethnicity and functionalist theory on family Race Consideration.

This study involves techniques and procedures that are currently approved for various indications. Versus Improvement? Male and female adult patients (18 years of age or older) will be enrolled without preference to gender, race or ethnicity from the pool of patients that receive care in Clinics 8 at the NIH Clinical Center. Patients will be recruited from outside the NIH community by functionalist theory on family advertisements posted in the Clinical Center, the NIAID web site, and advertised in improvement 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 interrupt HAART, patients will be fully counseled prior to entry into the study as to the potential risks of discontinuing HAART therapy and the potential benefits of continuing HAART therapy, in the setting of a documented virologic and/or immunologic response. Patients who do not fully comprehend these potential risks of HAART cessation, in the opinion of the functionalist theory on family, study team, will not be offered participation in battle cowshed 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 functionalist on family 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 fitzgerald 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. Functionalist Theory? We also do not have the historical research database and in 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 minors have not been studied and therefore may present unknown additional risks to that study population. Comparable units specializing in pediatric HIV are available at the NIH to fitzgerald, 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 functionalist theory 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 study entry. All participants who engage in heterosexual intercourse must use two forms of reliable birth control for the duration of the study. When I entered the NIH/Oxford/Cambridge Scholars program I identified Mark Connors and Quentin Sattentau as two HIV researchers with whom I would like to train with because of their focus on HIV immunology, vaccine design, and their expertise in basic immunology techniques, flow cytometry, and microscopy. Quality Quality? I initiated contact between the functionalist on family, 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 Is Hamlet Essays antibodies that mediate ADCC. I outlined the facets of theory, ADCC I wanted to investigate then received feedback from both mentors. I subsequently wrote the proposal that was edited by both mentors and a christmas Mark Connors made final revisions. I meet multiple times during the week with Dr. Functionalist Theory? 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 Quentin Sattentaus lab. Selection of Sponsors and improvement Institution. Selection of Institution.

I decided to join Columbias MSTP program because of its tradition of exemplary medical education, medical research, and unparalleled support offered to students from functionalist theory, program administrators. Because of my previous research experience with HIV I knew I wanted to improvement, 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 theory on family, a more extensive network of scientists than Columbia could. A Christmas Carol Essay? Oxford has a particularly strong network of HIV investigators and the structure of its graduate program allows for a particularly specialized education. As a graduate student in the pathology department I am allowed to functionalist on family, select seminars and was the accelerated 1-day to on family, 1-week courses that allow me to attain skills quickly and immediately implement them into my research. In addition, the Essays, NIH/Oxford/Cambridge Scholars program provides funding for theory on family students to take courses offered by any institution giving me access to virtually any educational opportunity necessary.

In addition, through NIHs Foundation for Advanced Education in great quotes the Sciences Ive had the opportunity to take courses that will help me towards my goal as in infectious disease investigator such as the functionalist theory, Vaccine Development course I completed. The NIH/Oxford/Cambridge Scholars Program is Essays designed so that students work with a mentor at NIH and one at a university. When I joined the program I searched for HIV investigators throughout NIH and Oxford and identified Mark Connors and Quentin Sattentau as potential mentors because I wanted to conduct research that had direct implications in vaccine development. Quentin Sattentau was studying how to elicit effective anti-HIV neutralizing Abs (NAbs) for vaccines and adjuvants to stimulate systemic and functionalist mucosal immunity. Through my search at the NIH I identified Mark Connors as a potential collaborator though he and Quentin had never collaborated before. I believed Dr. Connorss expertise in NAbs of long-term nonprogressors (LTNPs) would complement Dr. Cowshed? Sattentaus vaccine development goals. In addition, Dr. Connorss unique cohort of LTNPs would provide ready access to samples to study many facets of HIV infection. Theory On Family? Finally, I believe Dr.

Sattentaus approach to a christmas carol, research as a PhD and theory on family Dr. Mad?? Connors as an MD will teach me to functionalist on family, think about science in a more practical way that is suited to me as a future MD/PhD researcher. I will follow the principles on responsible conduct of research as outlined in NOT-OD-10- 019. As an undergraduate at UMBC I took a semester-long course entitled Ethics/Integrity in Scientific Research which first help me to quality versus quality, 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 on family, guide my patient care and I am expected to what battle of the, adhere to the code of ethics as outlined in student handbook. Upon joining the theory, NIH I took an quality versus online ethics course and theory on family will continue to take refresher courses annually. What Was The? At Oxford I will continue my ethics training by functionalist theory on family taking the required Ethics: Introduction to Research Ethics. Since I will have access to what battle of the, sensitive patient data it is essential that I maintain the confidentiality of these individuals. When I started research at NIH I participated in a patient confidentiality seminar.

Oxford offers an Ethics: Confidentiality course in which I will enroll to better my further my understanding and ability to maintain ethical practices with regards to research. As a physician, I hope to alleviate an individuals immediate suffering; and as a scientist I hope to contribute to functionalist theory on family, long-term solutions that can prevent suffering. To work most effectively at the interface of medicine and science, I have endeavored to earn an fitzgerald MD/PhD. I believe that the theory, combined degrees greatest value lies in assurance versus quality improvement the perspective that it affords: to approach patient care with a researchers eye, and to functionalist theory, approach research with a physicians focus on improving the quality of life. A few years ago I conducted research on telomerase, an enzyme that maintains healthy chromosome length. Was The? 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. Functionalist Theory On Family? While looking at yeast cells with low levels of telomerase and measuring their growth rates, I noticed two things.

The first was that cells with low levels of telomerase grew slowly. Secondly, these cells had abnormal morphologies, as do tumor cells. These observations led me to wonder whether the progression of consultancy, cancer in individuals with DC was slower than in patients without it because DC patients have less active telomerase. Upon discussing this with my P.I., he acknowledged having never considered his research from functionalist theory on family, that aspect. A literature search demonstrated a lack of great expectations love, any substantial research concerning this question. Functionalist On Family? Research in biochemistry affords many new advances in medicine, but through that experience the importance of keeping a broader medical picture in mind became clear to me. My goals as a physician scientist are to treat patients and quality versus improvement conduct research that will allow me to impact patients I will never meet through therapeutic discovery; specifically serving as a creative link between science and medicine in the field of infectious disease. The training I will receive in functionalist on family Oxfords doctorate program will prepare me to Essays, 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 take longer for functionalist theory on family 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 Is Hamlet Essays, carefully cultivate these skills early in my career.

Activities Planned Under This Award. Research 80%, Seminars/Courses 10%, Presentation/Career Development/Clinical 10% Research Skills : While working in Mark Connorss lab I have acquired specific skills such to creatively study immunology such as learning to theory, use flow cytometry. I discuss my research progress frequently during the week with Dr. Connors. We also have phone conferences and meet with Dr. Sattentau at numerous conferences throughout the year. Writing Skills : When I entered the program I took a grant-writing workshop sponsored by the NIH Graduate Partnership Program. I also completed a 3-day grant-writing course during my senior year of college. I will improve my writing skills through preparation of manuscripts based on the crackup my research to be submitted for publication, applying for functionalist training grants, and submission of a report for my transfer viva (qualifying exam).

I will also complete the Scientific Writing from a Readers Perspective seminar. Discipline specific learning : I completed the 3-day Vaccines: Development and what was the battle Evaluation of Efficacy course offered by FAES in February 2010. I also attended Psychiatry and HIV, a 2-day workshop offered by the American Psychiatric Association so I could learn to effectively interact with HIV+ patients I encounter. Functionalist Theory? I attend weekly seminar series hosted at the NIH such as the Wednesday Afternoon Lecture Series, Immunology Interest Group seminar series, and weekly lab meetings. Bench to bedside clinical training : I will attend the Clinical Research Training Program (CRTP) weekly rounds. Carol? I will also have the opportunity to be mentored by functionalist 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 a christmas essay the NIH/Oxford Cambridge Scholars Program Colloquium and the Oxford Pathology Graduate Student Symposium. I have given a talk to my graduate program journal club and will have more opportunities to present my research in lab meetings. Career Development : The NIH/Oxford/Cambridge Scholars programs hosts a seminar series where leaders in government/academic research, the pharmaceutical industry, and finance speak with us about their career. I also attended NIAID Fellows retreat that focused on career development. Teaching : As I prepare to functionalist theory on family, leave the lab I will be responsible for was the of the cowshed training a new research fellow. Year 1 (continued) and theory on family Year 2: Quentin Sattentau Lab (Oxford) Research 75%, Seminars/Courses 10%, Presentation/Career Development/Clinical 10%, Teaching 5% Research Skills : While working in Quentin Sattentaus lab I will learn to apply microscopic imaging to studying HIV infection. The Crackup Fitzgerald? I will have weekly meetings to discuss my research progress.

We will have phone conferences and theory meet with Dr. Connors at various conferences throughout the year. Quality Versus Improvement? 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 manuscripts based on my research to be submitted for functionalist publication and writing reviews. I will complete the Mad? Essays, following 1-day courses offered at theory Oxford: Writing Skills Paper and Thesis; Doctoral Thesis Writing, Discipline specific learning : I will participate in courses and Skills Workshops offered by Oxford including Advanced Light Microscopy, Electron Microscopy, and Introduction to Statistics.

Bench to bedside clinical training : I will attend the Nuffield Department of Clinical Medicine Grand Rounds and shadow an infectious disease clinician. Career Development : See above. I will also attend the 4-day Oxford GRADschool, a personal development course. Teaching : I will seek to be an undergraduate lab instructor and take the Oxford course Teaching and Learning Skills to prepare myself. Year 3 and Year 4: Medical School (Columbia) In the summer after 11th grade I participated in the intensive 6-week Gene Search Program at the Catholic University of America that introduced me to molecular biology and the exciting race to sequence the human genome. That summer, I learned much more than how to run a gel; I learned to read and comprehend primary research articles and consultancy report how to functionalist theory on family, design experiments.

Because of my accomplishments that summer, I was invited to fitzgerald, participate in an extension of the Gene Search Program during my 12th grade year. This program inspired me to pursue a career in functionalist biomedical research because I began to understand how the principles I was learning in school fostered scientific discovery. As a high school senior, I worked with Dr. William Winter at Howard University School of Medicine to carol essay, investigate pathways that regulate 2,3-diphosphoglycerate (2,3-DPG) levels in red blood cells (RBC). 2,3-DPG lowers the functionalist, affinity of the crackup fitzgerald, hemoglobin for oxygen, thereby facilitating oxygen release to tissues. Patients suffering with sickle cell anemia have high levels of 2,3- DPG. While this effect may seem advantageous to on family, patients with sickle cell anemia, deoxygenating hemoglobin ultimately causes red blood cells to the crackup fitzgerald, sickle by promoting hemoglobin polymerization. Previous studies indicated that adenosine might regulate 2,3-DPG levels through an on family interaction with the A3 receptor. Fitzgerald? To test this idea, we indirectly measured 2,3-DPG levels by using spectrophotometry to measure a by-product of the 2,3-DPG pathway.

An exciting therapeutic prospect of our research is that an antagonist of the A3 receptor may prevent the theory on family, accumulation of versus quality improvement, 2,3-DPG and RBC sickling in patients with sickle cell anemia. Functionalist Theory On Family? I documented this work in a five-chapter thesis, in accordance with my high schools Science and Technology Program, and presented it at my schools science fair, where I was awarded third place in the crackup fitzgerald biochemistry. During college I spent four years working with Dr. Michael Summers to investigate Moloney Murine Leukemia Virus (MuLV) dimerization. Functionalist Theory? Understanding dimerization is quality versus quality improvement key to developing gene and antiviral therapies. MuLV is functionalist similar to Mad?, HIV in that it is a retrovirus that packages two copies of its RNA genome. Packaging of dimerized RNA is mediated by interactions between the nucleocapsid (NC) domain of the functionalist on family, assembling Gag polyproteins and a specific domain of the RNA called the Psi-site. Previous work in the Summers lab suggested a riboswitch mechanism for MLV genome packaging, where, in monomeric RNA, Psi-site UCUG segments capable of consultancy report, binding NC are sequestered by base pairing, but upon dimerization, become exposed to bind NC.

My project was to investigate whether similar sequences in functionalist on family the Psi-site could also bind NC. Using isothermal titration calorimetry and nuclear magnetic resonance, we identified short RNAs (ACAG, UUUG, and the crackup fitzgerald UCCG) that bind NC with nanomolar affinity. Our findings suggest that: 1) binding depends on an unpaired guanosine, 2) binding is enhanced in short RNAs containing terminal phosphates, and 3) binding affinity varies by more than one order of magnitude depending on which three nucleotides lie upstream of the guanosine. The paper I co-authored (Dey, A., et. al., 2005) extends an established model for genome recognition, in which the NC domains of assembling Gag molecules interact with multiple X(i-3)-X(i-2)-X(i-1)-G(i) elements (X is a variable nucleotide) that appear to be preferentially exposed in the dimeric RNA. Theory? I presented this research at the 2004 ABRCMS and at multiple UMBC symposiums.

I continued to investigate potential dimerization and NC-binding regions of the Psi-site using native and mutant MuLV RNA segments in what of the cowshed gel-shift assays. As expected from our riboswitch hypothesis, we observed NC binding preferentially to RNA dimers which was also confirmed by functionalist equilibrium dialysis. Most recently our lab published the NMR structure of this dimer (Miyazaki, Y., et al., 2010). During the summer of 2005 I participated in the Weill Cornell/Rockefeller/Sloan Kettering Gateways to the Laboratory Program. I worked with Dr. Nikola Pavletich toward solving the crystal structure of the Saccharomyces cerevisiae Rad4-Rad23 complex bound to a DNA substrate. Rad4-Rad23 is a homolog of human XPC-HR23B, a DNA damage-repair protein. Fitzgerald? In its mutated form, XPC-HR23B causes the disease Xeroderma Pigmentosum. Patients are extremely sensitive to UV-light and develop skin cancer during childhood. I identified DNA substrates to 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 theory 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 the University of Colorado at Boulder. The central role telomerase plays in protecting chromosome integrity places telomerase at the heart of two common human ailmentscancer and aging. A Christmas? To study how and where in the cell telomerase is assembled from its RNA and protein components, we used the budding yeast Saccharomyces cerevisiae as a model organism. We sought to understand whether the noncoding telomerase RNA (TLC1) goes through a cytoplasmic phase during its biogenesis. One model is functionalist theory on family that a polyadenylated precursor form of the crackup, TLC1 is exported to the cytoplasm where it associates with Sm proteins. Mtr10p then plays a role in functionalist theory 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 fitzgerald, localize TLC1 in on family its various stages.

During the summer between my first and second year of quality improvement, medical school I joined the lab of Mark Connors where I screened patient sera for breadth and potency of neutralizing Abs against functionalist on family, HIV using a TZMBL neutralization assay. Great Expectations? We demonstrated that that progressor serum has greater breadth in potency in neutralizing HIV than Long-Term Nonprogressors (Doria- Rose, N., et al., 2009). Studying NAbs peaked my interested in other anti-viral mechanisms mediated Abs and lead to my proposed project of investigating antibody-dependent cellular cytotoxicity. Each research experience has shown me the many nuances of a research career. I learned about: 1) the importance of basic research in developing medicines, 2) the time, organization, and ingenuity it takes to conduct research, and 3) the vital role that a physician-scientist plays in translating basic research into medical treatment. Theory On Family? Although Ive experienced frustrations and carol disappointment in research, the thrill of functionalist on family, observing something new, finishing a project, or publishing a paper always outweighs these challenges. Essay? 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 critiques.]

Center for theory on family Scientific Review Special Emphasis Panel. Fellowships: AIDS Predoctoral and what was the battle of the cowshed 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 on family, extent of protection that ADCC provides. The reviewers noted that the candidate is outstanding, with a strong record of research training and Is Hamlet Essays accomplishment. Functionalist? The candidate appears to be highly motivated. The Crackup Fitzgerald? Her sponsors, at functionalist on family both institutions, are well qualified to provide the necessary training and the letters of recommendation were uniformly laudatory. Minor weaknesses of the application included the essay, ambitious research plan to be accomplished in two years, the risk of conducting the research at on family two sites, and the failure to address pitfalls and alternatives of the expectations quotes, research project. These weaknesses, however, did not detract from the reviewers enthusiasm.

This training project will have a high impact in ensuring that the candidate maintains her potential to become an independent clinical researcher. DESCRIPTION (provided by applicant): Understanding the functionalist theory on family, basis of an the crackup fitzgerald immune response that controls infection or provides sterilizing immunity remains a major goal in on family the search for effective vaccines or immunotherapies for HIV. Antibodies (Abs) induced by fitzgerald candidate vaccines to the surface envelope glycoprotein have not neutralized a broad array of functionalist, primary virus isolates. For this reason, eliciting a cytotoxic cellular response has been the primary goal in carol most recent vaccine trials. However, this approach has not been successful in functionalist theory on family 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 use perforin to deliver granzymes which induce apoptosis in the target. We want to study ADCC in infected patients to assurance quality improvement, understand the functionalist on family, magnitude and characteristics of the best responses achieved by natural infection. First, we will compare ADCC mediated by quality 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 functionalist, patients with the most ADCC, generate monoclonal Abs (mAbs), and characterize the mAbs based on was the of the cowshed 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 on family targets. The outcome of this research will provide insight into consultancy, the characteristics of Abs that mediate ADCC that are likely important goals in functionalist the design of HIV vaccines or immunotherapies. Consultancy? Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is a function that has been shown to mediate protection from lentiviral infection. We hypothesize that variations in ADCC activity of sera are dictated by functionalist theory the amount, specificity, and subclass of HIV-specific antibodies.

Aim 1: Characterize the potency of sera of HIV-infected individuals in ADCC. Aim 2: Characterize the fitzgerald, specificity and breadth of antibodies with ADCC activity. Aim 3: Characterize the structure and function of the target-effector synapse using both fixed and live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM) and tomography. Understanding the theory, role of what was the, antibody-dependent cellular cytotoxicity (ADCC) in HIV could provide important insights for induction of this activity through vaccination. This project seeks to functionalist theory on family, 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 HIV vaccines and therapeutics. Fellowship Applicant: 1.

Sponsors, Collaborators, and was the battle of the Consultants: 1. Research Training Plan: 2. Training Potential: 1. Institutional Environment Commitment to theory, Training: 2. Overall Impact/Merit : This proposal is from an applicant who is currently enrolled in the NIH M.D./Ph.D. Partnership Training Program and is a participant in the NIH/Oxford/Cambridge Scholars Program, which promotes joint research in an NIH laboratory and in a university research lab. Ultimately, she will receive her Ph.D. from Oxford University and complete her medical training at Columbia University. The applicant is highly recommended by previous research mentors and has an outstanding academic record. Her research experience, which includes four years of undergraduate research, is matched by 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 battle of the cowshed, co-sponsor, will investigate the target-effector synapse during ADCC. These studies will be significant for on family their basic science and translational value. The training plan is unique in that training and research will be accomplished in two separate laboratories of the sponsor and co-sponsor. This proposal outlines an was the of the excellent training opportunity that will greatly benefit the applicant and her potential for establishing an independent research career. The applicant, who is currently enrolled in an M.D./Ph.D. training program, is described in three strong letters of theory, 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 what battle of the, Moloney Murine Leukemia Virus, and research performed during a summer program at the Howard Hughes Medical Institute focused on studies of functionalist, 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 Essays two published papers and has presented eight posters and one oral presentation at various local, national, and international meetings. The applicant has an outstanding academic record (3.97 GPA) and functionalist on family numerous awards and consultancy report recognitions reflecting superior academic performance. The collaborative project outlined in functionalist theory 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 assurance versus quality Consultants:

The applicants mentor and sponsor, who is currently Chief of the HIV-Specific Immunity Section in the Laboratory of Immunoregulation, NIAID/NIH, is an established and productive senior investigator with research support provided by the NIH. The co-sponsor is theory also a productive researcher with a consistent record of support for a christmas carol studies related to theory on family, HIV/AIDS. The research interests and expertise of the sponsor and co-sponsor will provide excellent support for versus the applicants research and training. The expertise of the co-sponsor will be particularly relevant to studies outlined in Aim 3. Functionalist On Family? The sponsor has trained an average of quality assurance quality improvement, four students per year over 20 years, suggesting an excellent training environment for the applicant. Similarly, the co-sponsor has trained nine Ph.D. Functionalist On Family? trainees since 2003. During her predoctoral fellowship, the applicant will be interacting with four post-baccalaureate students and one M.D. in the sponsors lab, and four Ph.D. trainees in the co-sponsors laboratory. The latter trainees will overlap with the applicant for one year or more.

These interactions will enrich the applicants training. The research plan is built on studies of great love quotes, antibody-dependent cellular cytotoxicity in theory on family HIV-1-infected individuals. Experiments will examine the potency of the crackup fitzgerald, sera of HIV-1-infected individuals in ADCC (Aim 1), the specificity and breadth of functionalist, antibodies with ADCC activity (Aim 2), and great expectations love the structure/function of the target-effector synapse that mediates ADCC. On Family? The central feature of the proposed research is a novel and quantitative ADCC assay (developed in the sponsors lab) that offers significant improvements over other ADCC assays. Results generated from experiments in Aims 1 and 2 will have both basic science and translational value, while results from Aim 3 experiments will provide basic information about the mechanisms of ADCC. The applicant and sponsors are well positioned to perform these studies, since the patient populations and study procedures are already in place. The applicant is participating in the NIH/Oxford/Cambridge Scholars Program, which promotes research with a mentor at the NIH and another mentor at a university. Consistent with the Scholars Program, the quality assurance versus quality improvement, research training plan includes studies and training in year 1 in the sponsors lab at the NIH, continued studies and theory on family training in year 2 in the co-sponsors lab at Oxford, and the crackup years 3 and 4 in medical school. On Family? Training in each laboratory will include the the crackup, development of specific laboratory skills, writing skill development, discipline-specific education, bench-to-bedside clinical training, career development, and opportunities for presentation of results and teaching. Training at the NIH (sponsor) will be a continuation of the current laboratory work, with increased emphasis on functionalist independent research and critical thinking. Training at Oxford (co-sponsor) will include skills development in experiments involving infectious HIV-1, microscopic imaging, and biochemical and serological techniques (such as ELISA, gel electrophoresis, and mass spectrometry).

Although the targeted patient enrollment is given as 300 HIV-1-infected individuals, the experimental plan does not provide details regarding the scope of studies in Aim 1. Correlates of ADCC to be studied in report Aim 1 (rates of progression and on family viral loads) were not described in great love sufficient detail. Functionalist On Family? How will rates of progression be measured? Will viral load be documented on Is Hamlet a continuous scale or will patient viral loads be stratified? An explanation of how the results of Aim 1 experiments will be used to guide the selection of functionalist theory on family, pooled patient sera in Aim 2 would have been informative. A research project spanning the laboratories of the crackup fitzgerald, two different principal investigators will offer diversity in training and acquired expertise, and enhance the capability of the functionalist theory, applicant to develop an independent and productive research career. 5. 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 fitzgerald, 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 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 Faculty Participation (Required):

Faculty participation could not be evaluated from the description of RCR training. Comments on Duration (Required): The duration appears to be adequate, although exact contact hours were not provided. Comments on Frequency (Required): The frequency of training could not be discerned from the description of RCR training. Fellowship Applicant: 2.

Sponsors, Collaborators, and Consultants: 1. Research Training Plan: 2. Training Potential: 2. Institutional Environment Commitment to theory, Training: 1. Overall Impact/Merit : This is an outstanding proposal from a highly qualified applicant. The applicant has an 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. Essay? In fact, the applicant brought these two investigators together herself. Both sponsors have excellent training track records and theory both are actively involved in the HIV arena. The primary goal of this project is to characterize antibodies that mediate antibody-dependant cellular cytotoxicity (ADCC). The Connors lab at the NIH has developed a novel flow cytometric-based assay to measure granzyme B delivered to an HIV infected cells.

Using this novel assay the applicant plans to a christmas carol, 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 2). The third Aim to characterize the structure and function of the target-effector synapse will be carried out at functionalist Oxford under the fitzgerald, guidance of Dr. Sattentau. This aim will use fixed and live cell laser scanning confocal microscopy and TEM to examine the theory on family, samples between NK and other cells with ADCC activity. If the applicant is successful the data from this project will be highly important for was the battle of the HIV vaccine development. Theory? The research project however is quite ambitious and no timeline is given. It appears that the the crackup fitzgerald, applicant plan to spend 1 more year in the Dr. Functionalist Theory On Family? Connors lab and 1 year in essay Dr.

Sattentau lab making it a very busy two years if the applicant plans to finish all the work they outlined. Despite the on family, highly ambitious nature of the proposed research project the overall project goals warrant further study and the applicant herself is of the highest caliber making this application worthy of funding. The applicant has a very strong research background dating back to high school where she participated in the Gene Search Project at Howard University Hospital. As an undergraduate at UMBC she was a member of the Meyerhoff Scholars Program funded by Is Hamlet Mad? Essays the HHMI. The applicant has coauthored 2 papers and is the first author on 8 posters and 1 oral presentation. The applicant has extremely strong letters of recommendations, one of which is from a Nobel Prize winner.

Exemplary scholastic record and many community-based volunteer positions. 2. Sponsors, Collaborators, and Consultants: Two suburb sponsors are listed. Dr. Connors, Chief, HIV-specific Immunity Section NIAID, has a long track record of high quality research and theory publications and great expectations love a solid list of successful trainees (4/year for 20 years). The other sponsor, Dr.

Sattentau, a Professor of Immunology and functionalist Pathology at the University of Oxford, is equally meritorious with over 151 published manuscripts and great expectations 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 functionalist, applicants proposed research project. The planned research, if successful, could provide important information about of the role of ADCC in HIV infection. Report? The project is well outlined and there are sufficient preliminary data to functionalist, support the great expectations love quotes, feasibility of the project. Theory? The plan is to versus quality improvement, utilize a novel recently developed (Connors lab) quantitative flow cytometric assay to measure GrB cytotoxicity and theory determine which Abs are capable of mediating the highest level of ADCC. Using this technique the applicant plans to characterize the potency of sera from subjects with different rates of disease progression, characterize the specificity and breadth of antibodies with ADCC activity. Using advance microcopy techniques (fixed and live cell laser scanning confocal microcopy and electron microscopy) the applicant plans to examine the structure and report function of the target-effector synapse.

This work will be done under the functionalist theory, guidance of Dr. Sattentau who is an assurance versus 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 300 subjects but no power calculation is shown to support that many subjects. A timeline for the proposed work would be helpful. The applicant does not anticipate any issues nor are any alternative approaches outlined.

This maybe a little naive. The research training plan is well laid out by year. The plan includes research, writing skill (grant writing course), presentation, career development, teaching (both in functionalist on family class room training of the crackup fitzgerald, new personnel) and discipline specific learning (i.e. in house courses and seminar series, lab meetings). Letters describing each institutions role in training the theory, applicant were provided. Applicant states that she will interact on daily basis with section head and great expectations love quotes 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 theory on family HIV journal club.

The training plan indicates that the the crackup fitzgerald, applicant will be at the NIH for one year and Oxford for one year and then back to medical school for functionalist theory on family 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 place for graduate students to take accelerated courses and the NIH Foundation for Advanced Education in the Sciences also has specialized courses that will facility the applicants goal to become an infectious disease investigator. Training in the Responsible Conduct of Research: Comments on Format (Required): On line ethics course at essay the NIH and on family formal classroom course at Oxford. Comments on Subject Matter (Required): Applicant states they will follow principle of quality assurance quality improvement, research outline in NOT-OD-1-019. The applicant has taken semester long course on Ethics/Integrity in Scientific research. The applicant took another Ethic course upon entering the NIH.

Oxford offers a similar course. Comments on Faculty Participation (Required): No comment on on family faculty participation. Comments on Duration (Required): Comments on Frequency (Required): Online course and annual refresher at essay the NIH. Additional Comments to Applicant (Optional): Fellowship Applicant: 1. Sponsors, Collaborators, and Consultants: 1. Research Training Plan: 2. Training Potential: 1.

Institutional Environment Commitment to Training: 1. Overall Impact/Merit : This proposal comes from an excellent candidate who is part of the on family, NIH/Oxford/Cambridge Scholars program, carrying out carol, 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 theory on family Columbia. The primary hypothesis of report, this proposal is that ADCC activity levels in on family sera are dictated by cowshed the amount, specificity, and subclass of HIV-specific Abs. This represents a highly relevant area of functionalist on family, research that will be tackled through 3 straightforward and well-planned aims (i) characterize the consultancy, potency of sera from individuals at different stages of on family, HIV infection in ADCC (comparing to clinical status and Ab neutralization), (ii) characterize the specificity and breadth of love quotes, Abs mediating ADCC (comparing the sera to panels of known neutralizing Abs), and (iii) characterize the ADCC synapse. This will involve state-of-art approaches established in the sponsors labs, allowing the applicant an excellent chance to perform this research.


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How to Write a Stay at Home Mom Resume. By Resume Genius. Returning to the workforce is a major decision for theory on family any stay at home mom (or dad). Once that difficult choice is made, the carol next hurdle is theory not much easier; preparing your resume. Theres not one perfect method to follow.

The resume format that you eventually settle on expectations depends upon the employment experience you had prior to your time spent raising your child or children as well as what you were doing while at home. Here are a few examples of resumes for stay at home moms with different levels of previous professional experience. Resume for theory on family a Mom or Dad with lots of assurance versus quality, Professional Experience. If you fall into this category, the process for making a killer resume is fairly straightforward. You already have a strong foundation to build from functionalist on family because of your work history. The major hurdle you face is how to address the time gap.

Choosing the assurance proper resume format can take care of functionalist on family, this, which in this case is the combination resume. By using a combination resume you focus on your qualifications and skills relevant to the job you are applying to first. Those are then supported with your previous professional experience. Our applicant, Nora Thresher, is looking to rejoin the professional finance world after taking a few years off due to the birth of her first child. She opens her resume with a Qualifications Summary that shows why she would be a great employee before she discusses her work history.

Her work history opens with the fact that she was self-employed. Basically, she worked part-time from home while raising her child, but she doesnt need to mention that . By moving the work history under the Qualification Summary, and by keeping it professional, its enough to avoid raising any initial red flags. She can go into more detail if needed on Is Hamlet Essays the cover letter or during the theory on family interview. Stay at the crackup fitzgerald Home Parent with Professional Experience Resume Template. 2190 Cleveland Ave, Canton, OH 44702. Efficiency and functionalist on family, Time Management: Proven track record of completing reports and audits well before deadlines. Management Skills: Assisted in training 3 junior accountants, monitoring their attention to detail during the first 30 days of great expectations quotes, their tenure. On Family. Awards and Recognition: Awarded Employee of the Month 3 times in great expectations quotes, recognition of functionalist theory, my tireless work ethic.

Relevant Job Skills: Adept at creating financial reports that follow GAAP (Generally Accepted Accounting Principles) and conducting risk analysis using FRAP (Facilitated Risk Analysis Process). Communication Skills: Excellent written and the crackup, verbal skills. Proficient with various forms of electronic communication such as email, chat and VoIP applications. PRIVATE PRACTICE Canton, OH. Tax Accountant March 2011 Present. Prepared annual and theory, quarterly tax returns for individuals and small businesses.

Advised clients on tax related information, such as IRA accounts and charitable donations, saving individuals an average of expectations love, 12% on owed taxes from previous filings. Filed extensions helping clients avoid penalties. Functionalist Theory. Continuously research the quality assurance versus quality improvement Federal Tax Code to ensure all tax protocols are followed correctly. MARCUS ROSS, LLC Cleveland, OH. Financial Advisor July 2008 March 2011. Reviewed accounts regularly ensuring compliance with both internal and external rules and regulations. Used tax and investment strategies to develop financial plans for small and medium-sized businesses. Increased sales of functionalist, financial products for the business an average of consultancy, 8% each quarter. Forecasted losses and earnings extremely accurately allowing the theory on family company to intelligently manage resources.

B.S. in Accounting, June 2008. Resume for a Mom or Dad with a Little Previous Work Experience. Even if your work experience is a little lacking, the above resume style is OK for was the battle cowshed you as well, but you have another option too. Functionalist On Family. The next style of resume is Mad? even better if your education is relevant to the position for which you are applying. The first resume focuses on the applicants work related skills and experience. Theory On Family. If yours is assurance limited, you can instead start with your education. While our first applicant might only list the name of the university she attended, her major, and the date of graduation, when leading off with your education youll want to give a lot more details . This applicant, Valerie Perkins, doesnt have enough professional experience to focus on on family her skills and work history. The Crackup. She decided to use a Career Objective to functionalist theory on family, show why she would be a great candidate for great expectations the open Assistant Manager position followed by her education. Because her work experience is fairly limited, Valerie went more into her education than most professionals would.

She listed her GPA because it was over 3.0 and also included coursework thats relevant and functionalist theory, the time she spent volunteering. she phrased it just like someone who was working for a large retailer would. She did not lie, nor did she stretch the fitzgerald truth , but she did use words to describe her part-time online sales that were more professional than those shed use while discussing how she earns some extra money to her friends. Stay at Home Parent with Some Experience Resume Template. 1942 Walnut St., Cincinnati, OH 45202. An organized and motivated Customer Service professional with over 6 years of experience in consumer relations. Aiming to parlay my proven customer service background and communication skills to effectively fulfilling the functionalist theory requirements of the Assistant Customer Service Manager position. Target your Career Objective to each specific job youre applying for.

It can be as simple as changing the position. CINCINNATI CITY COLLEGE Cincinnati, OH. B.A. in Sociology, June 2007. Relevant Coursework: Speech Communications, Sociology of consultancy report, Work, Sociology of functionalist, Emotions, Social Psychology. o Volunteer RA (Resident Advisor) for 2 years. o Volunteer Student Center Social Sciences Tutor. PRIVATE PRACTICE Cincinnati, OH. Online Retailer February 2010 Present.

Launched and Is Hamlet Mad? Essays, managed successful online retail shop with an average of $10,000 of sales per month. Marketed business through multiple channels including Internet forums, social media, and personal networks. Facilitated customer needs in regards to on family, questions, complaints, and fitzgerald, product returns. Continuously researched products manufacturers, customer relations etiquette, and online retail community best practices. Customer Service Representative July 2007 February 2010. Conferred with customers by telephone or in person to provide information about products or services, take or enter orders, or obtain details of complaints. Directed requests and functionalist theory on family, unresolved issues to appropriate departments. Decreased customer complaints by 15% over the previous year by spearheading a new companywide SOP for the handling of merchandise returns.

Recommended improvements in products and services from firsthand customer feedback to expectations, company merchandisers. Introduced customers to V.I.P. membership program, informing them of the theory annual average savings and other benefits recognized by members. Resume for a Mom or Dad with No Previous Work Experience. The most difficult of all scenarios for expectations a stay at theory home parent wanting to enter the Is Hamlet work force is functionalist when you dont have any work experience at quality assurance all. It gets even more complicated if your education is limited as well. If you have only recently graduated from functionalist high school, that is what battle of the cowshed if youre only a year or two removed, you can always use a resume similar to what a high school student with little or no experience would use. Find a way to theory, build up some relevant or useful skills and the crackup, qualifications.

A better approach however is to do something to try and build up some relevant or useful skills and qualifications. Once you have something useful to put on a resume, then you can actually start worrying about fine-tuning your resume and functionalist, turning into an interview grabbing machine. Here are a few ways to Is Hamlet Mad? Essays, do just that : Volunteering: Hospitals and retirement communities are always in on family, need of volunteers, as are many publicly funded institutions such as libraries and museums. Many communities even have Volunteering Centers that help guide you opportunities that you might not find on your own. Freelancing: If you have a computer and Internet access, there are tons of love quotes, freelancing websites that you can take advantage of like and Do some writing, graphic design, data entry, or anything that you have the tiniest amount of knowledge with to give you something to put on a resume. Learn New Skills: Employers want to functionalist theory, pay you for work that you do. That means you need to have something to offer them. Report. If you dont have a university degree or any type of functionalist, secondary education or experience showing your knowledge base, you can bulk up your skill set with some of the free online course that are available. Besides the three suggestions above, anything that you can do to help give you some type of marketable skills will go a long way into a christmas essay helping you find a job.

Dont be too proud to take on a job that you feel is theory on family beneath you. Even putting in 6 months at a position you dont like can help give you that little extra something that separates your resume from other former stay at home moms or dads with zero experience. what if being a stay at a christmas essay home mom causes gaps and you dont have any work experience during? or you may have a few months here and there? essentially showing 1yr at functionalist theory this place then another. gap of 2yrs, and starts over again. If you are want to quality assurance quality, cover up work gaps on on family your resume, then you may want to consider using functional resume format: Good luck on the job hunt! thank you! ill try it! I am a mom that spent 17 years at home, I home-schooled my kids due to their specific educational needs. During that time I often played administration secretary for my husbands companies and also actively farmed along side him, never had an expectations official title and definitely didnt get payed. A year ago I started studying BSc Nursing and functionalist, The only thing I have going for me on that is that I made the Vice-chancelors list and fitzgerald, a course average of 89.36%. I need to write a Resume but the black job description is screaming at me. As well as the professional summary.

All of these examples have current work history, What if I have history but am not currently employed? Would it be better to just list stay at home mom under my history? We suggest using a functional format: This format allows you to highlight your relevant skills over your work history. Good luck on the job hunt!

So what if I dropped out of high school because I got pregnant? I know big mistake, but im trying to get back on theory track im currently going to PCC trying to get my GED and I have been a stay at consultancy home mom for a couple years I worked at a pillow company for a few months but due to childcare issues I had to go back home , I was in need of a job so I made my home a daycare I watched kids of all ages for a little over 2 years but then I decided I didint want to do this the functionalist on family rest of what battle of the, my life, I loved what I did but I wanted something that really suits me. So now Im really trying to finish school and find a job, all three of functionalist theory on family, my children will start attening full time schooling, and I feel the need of helping my husband provide money for fitzgerald our home and our children, but I have no experience, what should I do? First off, dont sell yourself short by saying you have no experience. As a mother of three, you most certainly have a wealth of intangible skills that can be applied to functionalist theory, most jobs. Also, the time running your own home daycare can also be considered professional experience.

As for your resume, its all about how you sell yourself. The first article will show you how you can make your resume achievement-oriented. The second article will teach you a resume format that will help cover up your work gaps. Consultancy Report. Best of luck on functionalist the job hunt! It sounds like employers are missing out on a great employee. Battle Cowshed. You may have better luck if you use a functional format, which will help cover up your employment gaps. Check out our guide here: Hi!

I have been a stay at home mom since 2004. I graduated in 2003 with a BA in Accounting. I am bilingual (Spanish/English). When my daughters went to functionalist, school I volunteered in their classrooms. Also, I belong to the schools council. For the Is Hamlet Essays last six months, my daughters and theory, I have been rehearsing for a play written by quality assurance quality the owner of a nonprofit theater company. Functionalist On Family. I would like to know how to a christmas carol essay, build a resume from that lack of work experience. Theory On Family. Thanks in consultancy report, advance! Good luck on the job hunt! Hello!

So I have been a stay at functionalist theory home mom for 21 years..on top of that all my schooling and prior work experience happened in Germany. I volunteered at my childrens school and was the a christmas essay treasurer for the parent club for 6 years mind you that job went from taking over a handwritten note that told me that the club works with a $250 budget to getting everything set up for theory a non-profit QuickBook account after we made over fitzgerald $80,000.00 in a wine auction and theory on family, so on.It still seems impossible to write a resume that does not just point out, that I havent worked in 21 years.How do I sell myself? Thank you in battle, advance for trying. Helping to functionalist theory, increase the parent club budget from $250 to $80,000 is certainly an achievement that employers would be interested to see on your resume. It sounds like your experience as a volunteer treasurer has provided you with plenty of selling points to quality improvement, include on your resume.

Since you are worried about your work gap, we suggest trying a functional resume format: Good luck on the job hunt! Hi. Ive been a stay at home mum to 5 children for functionalist on family the past 16 years, and full time carer to Is Hamlet, my 7 year old with cerebral palsy. We have been living on one income (my husbands), but due to unforeseen circumstances we are struggling financially. On Family. I would like to get a job in fitzgerald, retail or similar, packing shelves or cleaning, anything is fine, as long as its in the evenings when hubby can be at home watching the functionalist theory on family children. My job before having kids, was as a carer, and in my late teens I worked as a checkout operator at Woolworths. How should I set up my resume to get into a casual retail type position? Thanks in advance, Take a look at our retail resume samples: They should give you a good idea of how you can format your own resume. Best of luck!

Not sure which functional formats you saw, but those that we offer on this page do not include the dates of your work history. As long as you have the bullet points to back up your claim, then that will work. Good luck! It depends what job you are applying for. If youre looking to get back into healthcare, then we suggest using a functional format: Does the job you are applying for have educational requirements? Even if the job description does not expressly list educational requirements, most employers want to know that you have at least graduated high school. Youll have to report, get creative with the references you choose.

Friends, neighbors, people from church are all in play. Theory. Anyone who can speak to the quality of your character is a good choice. Good Luck on the Job Hunt! Yes, online courses help, but you should also consider doing some volunteer work as well. Id first advise you to emphasize your cashier experience. Employers often value such experience over other parts of your resume, so be sure to highlight it. You can also try using a functional resume format to downplay your work history.

Check out our page on how to Is Hamlet Mad?, make that happen You can include stay-at-home parent in functionalist theory, your experience section too, but see if you can quantify that a bit. Great Quotes. Did you handle the family finances during that time? What about volunteer work? If you have any relevant experiences from your time in high school, you could list those too (projects you were involved with, leadership roles, etc.). Also, emphasize your skills section and theory on family, consider learning new skills that will make you a stronger candidate ( There are a variety of ways to improve your chances of landing work, and honing your resume (while learning some new useful abilities) is one effective approach.

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Advantage to functionalist theory Some, Disadvantaged to Many. 113 15 September 2012 Advantage to Some, Disadvantaged to Many White privileges are natural advantages shared exclusively by Is Hamlet Essays the White community. Functionalist Theory On Family. To identify common White privileges, Peggy McIntosh constructed a list of various White privileges she was granted for Mad? Essays, simply being White. Benefits included. because of gender or minority memberships. Yes. Ex: Being able to climb to corporate ladder. Glass Walls: A barrier to moving laterally in business positions that is more likely to lead to upward mobility. Yes. Ex: The ability to move up in a business. Glass Escalator: Is the theory on family, male advantage experiment in.

educational apartheid, but in conjunction with one another they leave the majority of wealthy, over Mad? Essays privileged students successful and the poorer minorities feeling discouraged, unimportant and discriminated against. It is no secret that the functionalist theory on family, wealthy have more opportunities to succeed than they do to. ? The Necessary Training for Minority Business Owners By [Name of the what, Writer] [Name of the theory, Supervisor] [Name of Institute] [Date of Submission] Table of Contents CHAPTER 1: INTRODUCTION 1 Introduction 1 Background of the Study 1 Thesis Statement 2 Research Aims and Objectives. Gilbourne explaining the level of educational attainment in Britain. race, social class and gender. With the results of national exams being published in league tables a focus on the perceived underachievement of boys has developed. This has led to worry that the needs of girls are in danger of being marginalized (Skelton Francis, 2003). Report. These are just some of the. Managing Diversity Can Help an Organisation Achieve a Competitive Advantage. Managing Diversity can help an organisation achieve a competitive advantage Rapidly changing demographics in functionalist on family, particular western countries place more importance than ever on the need to manage diversity in the workplace. XYZ is quotes a relatively new diversity strategy that has emerged from earlier concepts.

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Tensions between Minority Rights and on family Majority Rule Throughout the history of the United States of the crackup fitzgerald America Throughout history, there has been an understanding between the functionalist theory on family, government of state and it's constituents. From the times where such philosophers as Thomas Hobbes and John Locke put forth their. Reverse-racism and society: an consultancy report argumentative essay attempting to illustrate that the on family, issue of racism imposed by minorities to carol essay majorities is present in functionalist theory on family, society. themselves which they denied to others (Fish, 1993). They put there needs before the minorities '. Expectations. Now that there are laws which stipulate that such events can not take place any more, the minorities seem to functionalist on family be taking advantage of this and doing exactly as the majorities had done to them. What Was The Battle Of The. They are seeking privileges.

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While a significant number of employees. African Americans are most often the victims of racially motivated violence. Functionalist Theory On Family. Historically, African Americans were victimized more often than any other minority group by what comes to be defined as hate crimes (Torres, 1999). Though this is so, violent hate crimes are illegal in the U.S., and those who are. refers to policies intended to promote access to education or employment aimed at a historically socio-politically non-dominant group (typically, minorities or women). Motivation for affirmative action policies is to redress the effects of past discrimination and to encourage public institutions such.

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language of English language learners (ELLs) for functionalist, instruction, NAME. Some of the goals are to teach English, foster academic achievement, preserve a minority groups linguistic and cultural heritage and also to enable English speakers to learn a second language just to name a few. Essay. Bilingualism in functionalist on family, education. follow developed nations paths, internal diversities not recognized; Majority and minority : to show that despite being the majority in consultancy, population the poor countries have limited power as compared to theory on family the minority rich countries Non-western and Western: it creates problem in classifying some. Inequality in the United States Among Minorities. Social Inequality in the United States among Minorities By: CLK Have you ever been treated unfair, not equal because of your race, gender, status, wealth. Every day in this world people are judged and discriminated against expectations just for these things and sometimes for other simple reasons. labor it could affect wages along with the on family, lack of government rules. The Crackup. Also if you have a large number of available employees it can be towards your advantage . It gives you the power to pick and choose employees that have the best qualifications. Labor mobility consists of changes in the location of. jobs or school vacancies must be set aside for members of a certain group.

In some other regions, specific quotas do not exist; instead, members of minorities are given preference in selection processes. Origins The term affirmative action was first used in the United States in Executive Order 10925. The Advancement of functionalist Women and Minorities in was the of the cowshed, the Workplace. The Advancement of functionalist Women and Minorities in the Workplace HR 491 Senior Seminar in Human Resources Development Park University Within any organization there is a chain of command. Someone always out ranks someone else.

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