Melissa Anderson - US grants

Anderson This longitudinal study of the doctoral experience links that experience to the nature of academic work and institutions. The fundamental question addressed in the study is: How do the experiences of doctoral students and early-career faculty reflect the nature of academic work, institutions, values and normative assumptions? This question is derived from the assumption that analysis of the doctoral experience and the transition into faculty life will contribute significantly to our understanding of academic work, the social environment of research and instruction, and the complex system of expectations, attitudes, beliefs, preferences, and norms that underlie the academic enterprise. The graduate experience provides a unique window into the normative bases of academic life as doctoral students experience it, and as they make decisions about the extent to which they will participate in an academic enterprise with those values. The project explores the assumptions, priorities, and preferences that shape the nature of academic work, by tracing their origins back to the experiences of doctoral students. The specific objectives of the proposed project are: 1) continuation of three years' previous data collection on the doctoral experience through a longitudinal series of interviews with approximately 100 of the 1994 cohort of doctoral students in 12 academic fields at a major research university; 2) extension of the interview series to a set of approximately 100 of the 1998 cohort of doctoral students in a variety of fields at the same university; 3) completion of an exploratory study of the outcomes of doctoral education and the transition from doctoral to postdoctoral life, through 30-40 telephone interviews with recent doctoral graduates; and 4) preparation of conference papers and publications based on the project.

animal tissue; proteins; enzymes; spectrometry; minerals; cardiovascular system; biomedical resource; biological products;

Reactive aldehydes derived from reducing sugars and lipid peroxidation play a critical role in the formation of advanced glycation end (AGE) products and oxidative tissue damage. We have recently proposed another mechanism for aldehyde generation at sites of inflammation that involves myeloperoxidase, a heme enzyme secreted by activated phagocytes. We now demonstrate that human neutrophils employ the myeloperoxidase-H2O2-chloride system to produce (-hydroxy and (,(-unsaturated aldehydes from hydroxy-amino acids in high yield. Identities of the aldehydes were established using mass spectrometry and high performance liquid chromatography. Activated neutrophils converted L-serine to glycolaldehyde, an (-hydroxyaldehyde which mediates protein cross-linking and formation of N(-(carboxymethyl)lysine, an AGE product. L-threonine was similarly oxidized to 2-hydroxypropanal and its dehydration product, acrolein, an extremely reactive (,(-unsaturated aldehyde which alkylates prote ins and nucleic acids. Aldehyde generation required neutrophil activation and a free hydroxy-amino acid; it was inhibited by catalase and heme poisons, implicating H2O2 and myeloperoxidase in the cellular reaction. Aldehyde production by purified myeloperoxidase required H2O2 and chloride, and was mimicked by reagent HOCl in the absence of enzyme, suggesting that the reaction pathway involves a chlorinated intermediate. Collectively, these results indicate that the myeloperoxidase-H2O2-chloride system of phagocytes converts free hydroxy-amino acids into highly reactive (-hydroxy and (,(-unsaturated aldehydes. The generation of glycolaldehyde, 2-hydroxypropanal and acrolein by activated phagocytes may thus play a role in AGE product formation and tissue damage at sites of inflammation.

DESCRIPTION (provided by applicant): This exploratory study will investigate challenges to the U.S. federal paradigm of research integrity in international research collaborations, as experienced and observed by NIH-supported scientists. Life-science research done outside the U.S. is subject to different standards of integrity oversight - some looser, some stricter, some simply different - and there is no international body or system by which problems and situations related to research integrity can be reviewed and addressed. The objective of this project is to provide empirical evidence on the variety, prevalence and impact of behaviors, situations and conditions in cross-national collaborations that either compromise or promote integrity in health research. The project is designed to extend three dimensions of current knowledge about integrity from the U.S. to the international context: a) the meaning and enactment of integrity in scientific research, b) its relationship to characteristics of research collaborations, and c) its relationship to characteristics of the broader research environment. Data will be collected through a series of 8 focus-group discussions with approximately 80 biomedical scientists who have experience in international research collaborations and 10 interviews with research administrators at top NIH-funded research universities. The focus groups and interviews will yield information about the range and forms of challenges to integrity that scientists have encountered or observed in cross-national research. Further data will be collected through a national survey of 6,000 NIH supported scientists. The survey will yield estimates of the prevalence and impacts of integrity issues in international collaborations, as well as assessments of the associations between research integrity and aspects of collaborations and the research environment. This research will investigate challenges to the U.S. federal paradigm of research integrity in international research collaborations, as experienced and observed by NIH-supported scientists. It will provide empirical evidence on the varieties and prevalence of behaviors, situations and conditions in cross-national collaborations that either compromise or promote integrity in health research. It will address the complications introduced by the expansion of international health-science research across countries with different interpretations and oversight of research integrity.

DESCRIPTION (provided by applicant): Rhesus monkey rhadinovirus (RRV), a gammaherpesvirus, is a close viral relative of Kaposi's sarcoma (KS)- associated herpesvirus (KSHV or HHV8), the causative agent of three human malignancies, Kaposi's Sarcoma (KS), primary effusion lymphoma, and multicentric Castleman's disease. RRV is an effective model to study the structure and assembly of gammaherpesviruses since it is able to produce high levels of progeny viruses in culture. The insights gained from studying mechanisms governing virally induced cancers can extend to a wide variety of malignancies, helping in the development of new treatments and therapies. Previous data from our laboratory have shown that RRV contains five gammaherpesvirus specific proteins in its tegument (middle) layer and of those, three have unknown functions, including the protein encoded by open reading frame 52, pORF52. Our hypothesis is that pORF52 plays a critical role in the structure of virions (viral particles) and their assembly as well as a potential role in cellular egress via a direct or indirect interaction with critical infrastructural components of the cell known as microtubules (MTs). Our methods of investigation include utilization of a wild-type (WT) RRV and an ORF52 deleted ( 52) RRV bacterial artificial chromosomes (BAC), each of which contains the remainder of the viral genome. Introducing these BACs into cells capable of supporting viral production will allow us to monitor the viral life cycle in the presence or absence of pORF52 production. The first goal will be to determine whether pORF52 is necessary for RRV DNA replication, a step preceding and required for progeny viral production. Quantitative analysis of the levels of individual viral proteins within the BAC containing cells will then help determine the requirement of pORF52 in the production of the various building block proteins necessary for viral assembly. In addition to these analyses, various imaging techniques will generate complementary data, helping us to examine the structure of the individual particles, the gross level of their production within the cells, as well as their intracellular localization and stage of maturation. In addition, media from cells containing either WT or 52 BAC will be assessed for a) infectious virus and b) production of structural viral proteins. Our preliminary results indicate that pORF52 is necessary for production of virus. To test this, 52 BAC containing cells will be complemented with the introduction of exogenous pORF52 and subsequently examined for the presence of infectious virions as described above. Additionally, RRV pORF52 expression by itself leads to its co-localization with, and stabilization of microtubules. Included in the second half of this proposal is a series of experiments that will investigate this potential interaction using microtubule binding assays, immunofluorescence and confocal microscopy, and assays that directly assess physical interactions between these two components. Combined with a series of mutations strategically introduced into pORF52, these approaches will allow the identification of the regions within the protein essential for not only its MT interactions but also its role in the viral life cycle. PUBLIC HEALTH RELEVANCE: The insights gained from studying mechanisms governing virally induced cancers can extend to a wide variety of malignancies, helping in the development of new treatments and therapies. In this application, we propose to characterize the actions of one specific protein in the life cycle of a subfamily of tumor forming viruses called gammaherpesviruses. We hypothesize that this particular protein may play an essential role in the formation of new viral particles and, thus, would be an attractive candidate for targeted anti-viral therapy.