This thesis explores the argument that inherent in medical treatment interventions for chronic hepatitis C, there are certain implicit ontological assumptions about the relationship between the body, disease, and society. Focusing primarily on biomedical practices, it is argued that these assumptions might have a profound effect on the world-views of patients undergoing them. This in turn, might have far-reaching sociological implications. Using a methodology specifically developed for the purpose of explicating the ontological assumptions inherent in medical treatment, the visual and narrative texts produced by thirteen hepatitis C positive participants are examined. A deconstructive analytical approach is then applied to these texts as they relate to the treatment interventions pursued by participants. An exploration of the way participants engage with, negotiate, and/or resist the discourses and assumptions inherent in biomedicine, traditional Chinese medicine, and to some extent naturopathy, is conducted. Two broad ways in which the participants visualise the relationship between disease and their bodies, relating to treatment undertaken, are identified. The possible social implications of these are then suggested. The first, and predominant view, is aligned with biomedicine. The relationship between disease and the body is antagonistic in this view. It is suggested that this way of seeing might naturalise xenophobic attitudes and perpetuate social conflict. The marginal view is related to non-biomedical treatments for hepatitis C. The relationship in this case is the result of a negotiated accommodation with the disease. It is suggested that such a view might allow for non-resistant social tolerance of that which is perceived of as new and different. This qualitative study contributes to the body of knowledge in the field of the sociology of health and illness in two ways: Firstly, it proposes a methodology that may be taken up or adapted for future sociological research, and secondly, it suggests something of the social and political nature of treatment decisions made by people living with chronic hepatitis C.

The separation, identification, and characterisation of the proteins present in a tissue or biological sample is called ‘proteomics’. This technique can be used for example to identify biomarkers and investigate signalling pathways. Increasingly, proteomics is being applied to the analysis of virus related samples; here two such examples are described. Presently there is no reliable non-invasive way of assessing liver fibrosis. Here a novel 2D-PAGE based proteomics study was used to identify potential fibrosis biomarkers. Serum from patients with varying degrees of hepatic scarring induced by infection with the hepatitis C virus (HCV) was analysed. Several proteins associated with liver scarring and/or viral infection were identified. The most prominent changes were observed when comparing serum samples from cirrhotic patients with healthy controls: Expression of inter-α-trypsin inhibitor heavy chain H4 fragments, α1 antichymotrypsin, apolipoprotein L1 (Apo L1), prealbumin and albumin was decreased in cirrhotic serum, whereas CD5 antigen like protein (CD5L) and β2 glycoprotein I (β2GPI) increased. In general, α2 macroglobulin (a2M) and immunoglobulin components increased with hepatic fibrosis whereas haptoglobin and complement components (C3, C4 and factor H-related protein 1) decreased. Novel proteins associated with HCV-induced fibrosis include the inter-alpha-trypsin inhibitor heavy chain H4 fragments, complement factor H-related protein 1, CD5L, Apo L1, β2GPI and the increase in thiolester cleaved products of a2M. The relationship between these changes is discussed. One of the accessory genes of the HIV viral genome encodes for the Nef protein. Nef is present in lipid rafts and increases viral replication within infected host cells by binding to a guanine nucleotide exchange factor, Vav. This leads to activation of a GTPase, Cdc42, however, the signalling pathway is poorly understood. 2D-PAGE based proteomics was used to identify differentially expressed raft-associated proteins by comparing T cells in the presence and absence of Nef. A ubiquitin conjugating enzyme UbcH7, which acts in conjugation with c-Cbl, was absent from the rafts of Nef-transfected cells. Vav ubiquitination was also absent from these rafts. In collaboration with Dr. Alison Simmons and Prof. Andrew McMichael the absence of UbcH7 in rafts was found to be caused by β-Pix forming a ternary complex with c-Cbl and activated Cdc42. Vav ubiquitination was restored and viral replication was diminished when β-Pix was knocked down providing a new candidate target for inhibiting HIV replication. This thesis demonstrates the use of proteomics in providing novel information for virus related samples. This influential technology benefits in both biomarker discovery to aid clinicians with early diagnosis of diseased individuals and in the elucidation of novel signalling pathways in infected cells to provide new candidate targets.

Hepatitis C Virus (HCV) is one of the most important causes of chronic liver disease, affecting more than 170 million people worldwide. The mechanisms of hepatitis C pathogenesis are unknown. Viral cytotoxicity and immune mediated mechanisms might play an important role in its pathogenesis. HCV infection and alcohol abuse frequently coexist and together lead to more rapid progression of liver disease, increasing the incidence and prevalence of cirrhosis and hepatocellular carcinoma. The cytopathic effect of HCV proteins, especially the core, E1 and E2 structural proteins, which induce liver steatosis, oxidative stress and cell transformation may be amplified by alcohol abuse. The purpose of this study was to characterize the liver damage mechanisms induced by HCV structural proteins and alcohol and to determine the potential molecular mechanism(s) that may promote chronic, progressive liver damage. A transgenic mouse model expressing HCV core, E1 and E2 was used to investigate whether alcohol increased HCV RNA expression. Real-time RT-PCR analysis of genes involved in lipid metabolism and transport confirmed their abnormal expression in the alcohol-fed transgenic mice. In addition, light and electron microscopy analysis were performed on liver tissues of transgenic mice on an alcoholic diet versus those on a normal diet, in order to identify histological changes. The severe hepatopathy in HCV transgenic mice was exacerbated by alcohol. Mitochondria and endoplasmic reticulum had severe abnormalities in the electron microscopy analysis. The second part of this study focused on adaptive immune responses, which may also play an important role in HCV pathogenesis. I focused my analysis on dendritic cells (DC), which have been the main suspects to explain immune impairment in HCV infection. Their powerful antigen-presenting function allows them to stimulate the antiviral response of CD4+ and CD8+ T cells, the effector cells of the immune system. This unique function of the DC makes them possible targets for immune evasion by the Hepatitis C virus. In this study, DCs were generated from mouse bone marrow cells. I investigated their maturation capacity in the presence of structural proteins of HCV. The impact of HCV core/E1/E2 polyprotein on DCs cytokine expression and ability to activate T-cell lymphocytes was also analyzed. A dysfunctional CD4 T cell response was observed after exposure of DCs to core/E1/E2 polyprotein, indicating inefficient CD4 priming, which might lead to chronic HCV infection in humans. The presence of the core/E1/E2 polyprotein reduced the DC maturation capacity and the expression of certain cytokines (IL-12, IFNg, IL-6, MCP-1) important for stimulation and chemotaxis of T cells and other immune cells. My studies contribute to the understanding of HCV pathogenesis and may have implications to the development of better therapies for HCV infection.

La infección por el virus de la hepatitis C (VHC) tiene gran importancia a nivel del trasplante hepático (HC) por varias razones. Por lo tanto, resulta de una gran importancia establecer el pronóstico post-trasplante en dichos casos.El objetivo del presente proyecto de tesis fue, por un lado, definir la historia natural de la hepatitis C postrasplante en nuestro medio; y por otro, establecer los factores de riesgo de evolución desfavorable. Dicha información es fundamental para validar la idoneidad de un trasplante hepático, así como para considerar la posibilidad de un tratamiento antivírico precoz y mejorar el manejo de la inmunosupresión de los pacientes.

Hepatitis C virus (HCV) replication is directed by NS5b, the viral RNA dependent RNA polymerase (RdRp). To date, our understanding of the HCV polymerase has come almost entirely from genotype 1. The aim of this study was to examine the influence of sequence variation in the polymerase region by characterising a polymerase derived from genotype 3a. The genotype 3a CB strain polymerase was cloned into the bacterial expression vector pTrcHis2C incorporating a hexahistidine tag to facilitate purification. An optimised process produced 2.5 mg of highly purified recombinant protein per litre of bacterial culture. The 3a preparation possessed an RdRp activity and could utilise both homopolymeric and heteropolymeric RNA templates. Optimal activity was seen at 30oC at pH 8 in reactions containing 160nM enzyme, 10??g/ml RNA template and 2.5mM MnCl2. Subsequently, three genotype 1b polymerases including the HCV-A, Con1 and JK1 strains were cloned for the comparison of activity under identical conditions. Steady state kinetic parameters for GMP incorporation revealed the 3a polymerase exhibited the highest activity, with an almost two fold higher catalytic efficiency (Kcat/Km) than HCVA-1b, primarily due to differences in Km for GTP (2.984??M vs 5.134??M). Furthermore, the 3a polymerase was 3.5 fold and 15 fold more active than JK1-1b and Con1-1b respectively. Improving our understanding of the influence of sequence difference on polymerase activity, particularly in the context of replication will be crucial to developing effective antiviral therapies.

Hepatitis C is known as the ??silent epidemic??. Globally 170 million people live with chronic hepatitis C, yet it receives little policy, media or public attention. In developed countries the blood-borne virus is primarily transmitted through illicit drug injecting practices, aiding its silenced and stigmatised status. In this thesis I uncover some of these silences by exploring the narratives of forty people living with hepatitis C in New Zealand and Australia. My status as a person living with hepatitis C informed all aspects of this research project; I therefore also include my own experiences, foregrounding researcher reflexivity and the co-constructed nature of the interview process. My aims are both practical and theoretical. On a practical level I explore the experiences of people living with hepatitis C in order to inform recommendations for policy, research and practice, while also working to elucidate and employ an approach that allows for an analysis of the ill body as a lived experiencing agent, located in a substantive web of connections whereby discourse, corporeality and sociality, inform and mediate one another. To this end I employ a ??political phenomenology?? influenced by phenomenological and poststructuralist theoretical approaches. The central, previously under-researched, issues that arose in participants?? narratives structure the chapter outline, with results chapters focusing on participants?? experiences of diagnosis, living with hepatitis C, stigma, support group membership, alcohol use, and hepatitis C treatment. For many participants, it was found that living with hepatitis C was a liminal experience where distinctions between what it was to be healthy or ill were not clear-cut. Indeed, many of the participants?? narratives exposed the inadequacy of Western binary categorisations to speak to their experiences of living with hepatitis C. Throughout this thesis it can be seen that the meanings that participants ascribed to health, illness, and their hepatitis C were fluid and contextual, informed by the interplay of corporeality and discourse. From this interplay comes the ability to speak into the gaps of dominant discourses, creating the potential for the disruption, or subtle realignment, of normative ways of knowing.

Hepatitis C is known as the ??silent epidemic??. Globally 170 million people live with chronic hepatitis C, yet it receives little policy, media or public attention. In developed countries the blood-borne virus is primarily transmitted through illicit drug injecting practices, aiding its silenced and stigmatised status. In this thesis I uncover some of these silences by exploring the narratives of forty people living with hepatitis C in New Zealand and Australia. My status as a person living with hepatitis C informed all aspects of this research project; I therefore also include my own experiences, foregrounding researcher reflexivity and the co-constructed nature of the interview process. My aims are both practical and theoretical. On a practical level I explore the experiences of people living with hepatitis C in order to inform recommendations for policy, research and practice, while also working to elucidate and employ an approach that allows for an analysis of the ill body as a lived experiencing agent, located in a substantive web of connections whereby discourse, corporeality and sociality, inform and mediate one another. To this end I employ a ??political phenomenology?? influenced by phenomenological and poststructuralist theoretical approaches. The central, previously under-researched, issues that arose in participants?? narratives structure the chapter outline, with results chapters focusing on participants?? experiences of diagnosis, living with hepatitis C, stigma, support group membership, alcohol use, and hepatitis C treatment. For many participants, it was found that living with hepatitis C was a liminal experience where distinctions between what it was to be healthy or ill were not clear-cut. Indeed, many of the participants?? narratives exposed the inadequacy of Western binary categorisations to speak to their experiences of living with hepatitis C. Throughout this thesis it can be seen that the meanings that participants ascribed to health, illness, and their hepatitis C were fluid and contextual, informed by the interplay of corporeality and discourse. From this interplay comes the ability to speak into the gaps of dominant discourses, creating the potential for the disruption, or subtle realignment, of normative ways of knowing.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 5 uppsatser.

Thesis (Ph. D.)--University of Washington, 1997. / Vita. Includes bibliographical references (leaves [42]-49).