O presente estudo avaliou materiais de 50 pacientes com cirrose hepática pelo vírus da hepatite C (VHC) que foram submetidos ao transplante hepático no Hospital das Clínicas da Faculdade de Medicina de São Paulo no período de 1993 a 2004, sendo divididos em dois grupos: Grupo 1 (33 pacientes com cirrose pelo VHC) e Grupo 2 (17 pacientes com cirrose pelo VHC com carcinoma hepatocelular). Nosso objetivo foi estudar a ocorrência da infecção oculta pelo VHB em pacientes com cirrose pelo VHC com ou sem CHC através do estudo molecular do genoma viral (DNA do VHB) no soro, tecido hepático tumoral e não tumoral pela utilização da técnica de Reação em Cadeia da Polimerase (PCR), pelos métodos in house e em tempo real. Todos os pacientes eram HBsAg negativos, possuíam soroteca e bloco de explante hepático em parafina, não apresentando concomitância com doenças hepáticas colestáticas, metabólicas e autoimunes. Foram avaliados os prontuários por um único pesquisador no sentido de coletar informações tais como: sexo, idade, dados de exames laboratoriais bioquímicos, sorológicos, ?fetoproteína e coagulação; além de dados clínicos tais como ascite e encefalopatia hepática para cálculos de índices prognósticos da cirrose (Child e MELD). Todo o material de explante hepático teve o Escore de Ishak e a Classificação das Sociedades Brasileiras de Patologia e Hepatologia para hepatites crônicas avaliados, assim como a Classificação de Edmondson e Steiner para os que apresentassem CHC. A técnica de PCR in house para detecção do DNA do VHB no soro e em tecido hepático tumoral e não tumoral apresentou negatividade em todas as amostras. Na técnica de PCR em tempo real apenas um caso do grupo 2 foi positivo no soro (sexo masculino, 66 anos, Anti-HBC total isolado e CHC); no tecido hepático tumoral no grupo 2 tivemos dois casos com resultados indeterminados e no tecido hepático não tumoral também do grupo 2, tivemos dois casos indeterminados. O grupo 1 não apresentou positividade para nenhuma das técnicas utilizadas. Concluímos que entre nossos pacientes com ou sem carcinoma hepatocelular associados à cirrose hepática pelo VHC, a infecção oculta pelo VHB foi muito baixa, provavelmente devido à baixa prevalência da infecção pelo VHB na nossa população / This study evaluated serum and liver tissue samples from 50 patients with liver cirrhosis due to hepatitis C virus (HVC) that underwent liver transplant at the Hospital das Clínicas - University of São Paulo School of Medicine during the period of 1993 to 2004, divided into two groups: Group 1 (33 cirrhotic patients due to HCV) and Group 2 (17 cirrhotic patients due to HCV with hepatocellular carcinoma - HCC). Our aim was to study the occurrence of occult HBV0 infection in cirrhotic patients due to HCV with or without HCC through the molecular study of HBV DNA in the serum, tumoral liver tissue and non tumoral liver tissue by the polymerase chain reaction (PCR) techniques using in house and real time PCR. All the patients were HBsAg negative, having previous serum samples frozen at -20ºC and liver tissue explanted in paraffin, without presenting concomitant cholestatic, metabolic and autoimmune liver diseases. The following variables were collected: gender, age, biochemical and coagulation laboratory tests and HBV serology (HBsAg, anti-HBc total, anti-HBs). Among the clinical data, ascites and encephalopathy were collected for the Child and MELD prognostic indexes. In the explanted liver tissue the Ishak\'s Score, The Brazilian Society of Pathology and Hepatology Classification for chronic hepatitis, and Edmondson and Steiner Classification for HCC were applied in the liver tissue. All samples with or without tumoral liver tissue and serum were negative for HBV DNA using in house PCR technique. By the real time PCR technique only one case from Group 2 was HBV DNA positive in serum (male, 66, isolated anti-HBc total positive and HCC). In the tumoral and non-tumoral liver tissues there were two indeterminated HBV DNA cases among Group 2 patients. All samples for Group 1 patients were negative for HBV DNA using both techniques. In conclusion, our study has shown the extremely low occult hepatitis B virus infection among the HCV cirrhotic patients with or without HCC, maybe due to the low HBV past infection among the Southeastern Brazilian population

Die systemische Autoimmunerkrankung Systemischer Lupus Erythematodes (SLE) ist durch die Produktion von autoreaktiven Antikörpern charakterisiert. In wie weit veränderte B-Zellrezeptor (BZR) Signalwege oder Co-Rezeptoren in diesem Prozess involviert sind, ist noch nicht ausreichend im humanen SLE untersucht worden. Aus diesem Grund wurde in der vorliegenden Arbeit eine detaillierte Analyse des inhibitorischen Co-Rezeptors CD22, der Kinase Syk und Akt in B-Zellen des peripheren Blutes von SLE Patienten durchgeführt. SLE Patienten zeigten eine Dysbalance in BZR abhängigen Signalwegen, welche eine B-Zellsubpopulationen unabhängige Reduktion der p-Syk/p-Akt Ratio versursacht. Diese Verschiebung könnte zu einer defekten negativen Selektion und somit zur Bildung von autoreaktiven Zellen führen, die wiederum durch Überlebensvorteile persistieren könnten. Zusätzlich wurde im peripheren Blut von SLE Patienten eine bislang nicht bekannte CD27 Syk++ B-Zellpopulation nachgewiesen. Diese wies, trotz des fehlenden Gedächtnismarkers CD27, Gedächtnismerkmale auf und könnte für die bekannte erhöhte Plasmazell-induktion in SLE Patienten verantwortlich sein. Somit konnte Syk als intrazellulärer Marker einer Gedächtnispopulation identifiziert werden. Des Weiterem stellt die Wiederherstellung der Balance von Syk- und Akt Phosphorylierung nach BZR Aktivierung einen erfolgsversprechenden Therapieansatz bei SLE Patienten dar, um die Entstehung und das Überleben von autoreaktiven B- und Plasmazellen besser kontrollieren zu können. / Systemic lupus erythematosus (SLE) is a severe systemic autoimmune disease in which loss of tolerance to nucleic acids results into the production of autoreactive antibodies (Ab) Therefore, B cells might play a key role in the pathogenesis of this disease. However, abnormalities of BCR associated co receptors and downstream kinases with potential implications in selection processes are rare for human SLE. Thus, a comprehensive analysis of the inhibitory BCR co-receptor CD22, the spleen tyrosine kinase (Syk) and the pro-survival serine kinase Akt has been undertaken to gain new insights into potential BCR signaling disturbances in this autoimmune disease. This data indicate that B cells from SLE patients display an intrinsically disturbed balance of BCR related signaling pathways, resulting in a B cell subset independent reduced p-Syk/p-Akt ratio. This may lead to a diminished BCR dependent negative selection and enhanced survival of SLE B cells, permitting the emergence of autoreactive B and plasma cells. Furthermore, SLE patients exhibit an increased frequency of a novel CD27-Syk++ B cell subset with memory features, enhanced tonic BCR signaling and the capacity to differentiate in auto-Ab secreting cells. The current study provides evidence that the use of intracellular markers, such as Syk, could permit a more precise delineation of CD27- memory B cell subsets in autoimmune diseases since the conventional used memory marker CD27 has some limitations. In addition, the balance between the BCR associated kinases Syk and Akt might be a promising therapeutic target to reduce the occurrence of autoreactive B and plasma cells.

Die b -Thalassämie gehört weltweit zu den häufigsten monogenen Erbkrankheiten. Die Thalassämien treten endemisch in der Bevölkerung des Mittelmeerraumes, in Westafrika und in weiten Teilen Asiens auf. In der einheimischen Bevölkerung der Bundesrepublik Deutschland gehört die homozygote Form der b -Thalassämie zu den seltenen Erkrankungen. Häufiger ist das Auftreten der heterozygoten Form, die als Differentialdiagnose der mikrozytären, hypochromen Anämie eine besondere Rolle spielt. Blutproben von 214 deutschen Personen mit einer heterozygoten b -Thalassämie wurden mittels Allel-spezifischer Oligonukleotid-Hybridisierung, Restriktionsanalyse und direkter Sequenzierung PCR-amplifizierter DNA analysiert. Insgesamt konnten 96,3 % (206/214) der Proben molekular charakterisiert werden. Die mediterranen Mutationen stellen einen Anteil von etwa 2/3 aller identifizierten Veränderungen, häufig sind insbesondere NS 39, IVS1-110 G ® A und IVS1-1 G ® A. Das übrige Mutationsspektrum setzt sich aus sehr seltenen Mutationen (IVS1-1 G ® T, IVS1-2 T ® G, IVS1-2 T ® C, NS 15 G ® A, NS 121 G ® T, FS 8/9 +G, FS 44 -C, FS 51 -C, FS 82/83 -G, Initiations-Kodon-Mutationen ATG ® ACG/ ® GTG/ ® ATA) und einer neuen Mutation (IVS1-129 A ® G) zusammen. In 6 Fällen konnte nach vollständiger molekularer Analyse kein Gendefekt als Ursache der b -Thalassämie gefunden werden. Diese Probanden könnten b -Thalassämiedeterminanten tragen, die nicht an den b -Globingen-Komplex gekoppelt sind oder regulative Sequenzen außerhalb des b -Globingens darstellen. Die erhobenen Daten zeigen, daß der Ursprung der b -Thalassämie in der deutschen Bevölkerung in den Mittelmeerländern liegt, ein Drittel der Fälle scheint sich jedoch lokal entwickelt zu haben. / The b -thalassemia belongs to the most common monogenic disorders worldwide. Endemically in the Mediterranean population, some parts of Asia and Western Africa, b -thalassemia is a rare disease in Germany. Nevertheless, heterozygous forms of b -thalassemia minor occur more frequently in the German population and should be considered in the differential diagnosis of hypochromic anemia. To investigate the molecular biological background of b -thalassemia in Germany, 214 non-immigrant German individuals suffering from heterozygous b -thalassemia were characterized by allele-specific oligonucleotid hybridization, restriction analysis and sequencing of the b -globin gene. By these techniques, 26 different mutations were identified. Most frequently, the Mediterranean mutations NS 39, IVS1-110 G ® A, and IVS1-1 G ® A were detected. Although otherwise rare, the frameshift mutation of codon 83 (FS 83 -G) was also relatively common (5 %) in the analyzed population. Other previously described mutations (IVS1-1 G ® T, IVS1-2 T ® G, IVS1-2 T ® C, NS 15 G ® A, NS 121 G ® T, FS 8/9 +G, FS 44 -C, FS 51 -C, initiation codon mutation ATG ® ACG/ ® GTG/ ® ATA) were demonstrated in < 10 individuals. Interestingly, sequence analysis identified a novel mutation affecting position -2 of the splice acceptor site (IVS1-129 A ® G). In 6 individuals diagnosed as heterozygous b -thalassemia, a mutation of the b -globin gene could not be demonstrated. The data indicate the b -thalassemia to be introduced from the Mediterranean population into Germans in two-thirds of the cases whereas the remaining third probably is of local origin.

Das FMC7-Antigen, eine unbekannte B-Zell-Membranstruktur, dient als eines der immunphänotypischen Grundkriterien zur Diagnose der typischen B-CLL. Die unterschiedliche CD20-Expressionsintensität ist auch ein charakteristisches Merkmal bei der Subtypisierung der Lymphomentitäten. In der vorliegenden Arbeit wurde die Modulation des CD20- und des FMC7-Antigens während einer Therapie mit CD20-Antikörper (Rituximab) bei Patienten mit indolenten B-Zell-Lymphomen untersucht. Bei der durchflusszytometrischen Untersuchung quantitativer und qualitativer Charakteristika der monoklonalen B-Zell-Population bei CLL- (n=12) und Non-CLL-Patienten (n=10) unmittelbar nach der Antikörperinfusion und bis zur 8. Woche nach der Therapie wurden parallele Veränderungen von FMC7 und CD20 festgestellt. Die Anzahl und die Fluoreszenzintensitäten der für die beiden Antigene positiven Zellen korrelierten signifikant sowohl bei der malignen Zell-Population (Kurzzeitbeobachtung: n=89; r=0,9; p / The FMC7, although being an unknown structure of the B-cell membrane, represents one of the basic immunophenotypic criteria for the diagnosis of the typical B-CLL. A different CD20 expression is a characteristic sign for the sub-typing of lymphomas also. The underlying study investigated the qualitative and quantitative modulation of the CD20 and FMC7 antigens in patients with indolent B-cell lymphomas (CLL, n=12 and non-CLL, n=10) during the therapy with a CD20 antibody (rituximab). Concomitant changes of FMC7 and CD20 expression were found immediately after rituximab infusion and up to 8 weeks thereafter. A correlation was seen for the number of positive malignant cells and for the corresponding fluorescence intensity (short-time observation n=89; r=0.9; p

Renal transplantation is the optimum treatment for end-stage renal failure. B cells have been identified in chronic allograft damage (CAD) and are associated with the development of tertiary lymphoid tissue within the human renal allograft. To investigate this pathology we utilized a mouse model of renal transplantation. A mouse model of kidney transplantation was first described in 1973. Although the mouse model is technically difficult it is attractive for several reasons: the mouse genome has been characterized and in many aspects is similar to man and there is a greater diversity of experimental reagents and techniques available for mouse studies than other experimental models. We reviewed the literature on all studies of mouse kidney transplantation to report the donor and recipient strain combinations that have been investigated and the resultant survival and histological outcomes. Some models of kidney transplantation have used the transplanted kidney as a life-supporting organ, however in many studies the recipient mouse’s native kidney has been left in situ. Several different combinations of inbred mouse strains have been reported, with varying degrees of injury, survival, or tolerance due to haplotype differences. Both cellular and humoral rejection processes have been observed. This model has been exceptionally useful as an investigational tool to understand multiple aspects of transplantation including acute rejection, cellular and humoral rejection mechanisms and their treatment. Furthermore this model has been used to investigate disease mechanisms beyond transplant rejection including intrinsic renal disease and infection-associated pathology. We performed renal transplantation in mice to model CAD and identified B cells forming tertiary lymphoid tissue with germinal centres. Intra-allograft B220+ B cells comprised of IgMhigh CD23- marginal zone, IgMlo CD23+ follicular zone and IgMlo CD23- transitional-type B cells similar to spleen, and these compartments had elevated expression of CD86. Depletion of B cells with anti-CD20 was associated with an improvement in CAD but only when administered after transplantation and not before. Isolated intra-allograft B cells were cultured and shown to synthesise multiple cytokines, the most abundant of these being GRO-α (CXCL1), RANTES (CCL5), IL-6 and MCP-1 (CCL2). Tubular loss was associated with T cell mediated injury and interstitial fibrosis, whilst type III collagen deposition driven by F4/80+ macrophages and PDGFR-β+ and transgelin+ fibroblasts, all of which were reduced by B cell depletion. In this report we show that intra-allograft B cells are key mediators of chronic damage to the transplant allograft kidney by cytokine orchestration of T cell, macrophage infiltration and fibroblast activation.

To date, masculine honour beliefs have been studied in the context of insults, threats and moral transgressions, and almost exclusively linked to aggressive emotions (e.g., anger) and behaviour (e.g., fights, confrontations). Here, it is proposed that masculine honour beliefs can also be associated with subtle, withdrawal-related behaviours, such as reluctance to engaging in feminine tasks and befriend feminine men. Furthermore, based on the theory suggesting that manifest indicators of a culture of masculine honour are expressions of individuals' overactive 'reputation maintenance psychology', I tested whether these subtle behaviours are underpinned by reputation maintenance concerns. Using self-report measures and different cultural samples (UK, Turkey, Saudi Arabia), the studies reported here as a whole provided evidence for the proposed associations and the reputation maintenance account. Studies 1a-b and 2a-b established an association between masculine honour ideals and men's self-presentations using masculine traits, as well as disfavourable judgments of effeminate men. Studies 3a-b and 4 focused on examining a voluntary relationship decision (choosing to associate oneself with a target as friends) to make reputational issues more salient and demonstrated that men who endorse higher levels of masculine honour beliefs were more reluctant to being friends with effeminate men. Study 4 further showed that this was due to high honour-endorsing men's concerns that being associated with an effeminate man who is perceived as lacking coalitional value would damage their own reputation among male friends. Focusing on the issue of men's disinterest in domestic roles such as child care, Studies 5a-b and 6 demonstrated a relationship between masculine honour beliefs and men's negative feelings (shame, frustration) about being a primary caregiver to their own children and revealed that this is due to high honour-endorsing men's concerns of losing reputation among their male friends, but not due to their wives' reduced appreciation of them. Taken together, these findings extend our understanding of individuals socialized with masculine honour norms, and also offer more nuanced explanations of men's anti-effeminacy bias and disinterest in communal roles.

INTRODUÇÃO: A infecção pelo vírus da hepatite B (VHB) é responsável por grande parte das doenças hepáticas crônicas em todo o mundo. Em pessoas vivendo com HIV/Aids (PVHA) a infecção pelo VHB tem maior risco de evolução para cirrose e carcinoma hepatocelular. A vacina da hepatite B é importante na prevenção de doença potencialmente grave, particularmente em PVHA, já que ambos os vírus têm as mesmas vias de transmissão e a coinfecção tem uma alta morbidade. Indivíduos imunocompetentes têm uma boa resposta humoral após uma primeira série de vacina da hepatite B e não há recomendações de rotina para doses de reforço. PVHA podem ter uma pior resposta à vacina da hepatite B, quando comparada à resposta em indivíduos imunocompetentes e a duração da imunidade nesses pacientes é desconhecida. OBJETIVOS: Geral: Avaliar os marcadores sorológicos de proteção e infecção pelo VHB em pacientes adultos vivendo com HIV/Aids, vacinados previamente para hepatite B. Específicos: Avaliar a persistência dos anticorpos anti-HBs em PVHA vacinadas previamente para hepatite B e que apresentaram resposta humoral protetora inicial; avaliar a resposta sorológica à revacinação para hepatite B nos pacientes vacinados previamente e que não apresentaram resposta humoral protetora inicial; investigar a presença de marcadores sorológicos de infecção pelo VHB em PVHA vacinadas previamente para hepatite B. MÉTODOS: Estudo observacional de coorte retrospectiva de PVHA vacinadas primariamente para hepatite B entre 2001 e 2002. Marcadores sorológicos de infecção e proteção para o vírus da hepatite B foram investigados nesses pacientes que ainda estavam em acompanhamento no Serviço de Extensão ao Atendimento de Pacientes HIV/Aids (SEAP), da divisão de Clínica de Moléstias Infecciosas e Parasitárias do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo entre 2012 e 2014. RESULTADOS: Uma coorte de 121 PVHA foi analisada quanto à soroconversão e persistência do anti-HBs. A maioria era do sexo feminino (54,5%) com média de idade de 50,1 anos. Destes pacientes, 58 (grupo 1) eram inicialmente respondedores à primeira série da vacina (anti- HBs >=10 mUI/mL) e 63 (grupo 2) eram não respondedores. Após um período mediano de avaliação de 11 anos, nenhum dos pacientes teve evidência sorológica de infecção pelo VHB e 41/58 (70.7%) dos inicialmente respondedores mantinham anti-HBs >= 10 mUI/mL. Maior contagem de células T CD4+ e anti-HBs >= 100 mUI/mL, no momento da primeira série vacinal, estiveram associados à persistência de anti-HBs. Durante o período avaliado, 35/63 (55.6%) dos pacientes inicialmente não respondedores (grupo 2) soroconverteram com sucesso (anti-HBs >= 10 mUI/mL) em resposta a uma ou mais doses de reforço vacinal. Foi associado à soroconversão do anti-HBs o número de doses de reforço recebidas. A partir do momento da soroconversão (anti-HBs >=10 mUI/mL), 70 pacientes não receberam nenhuma dose adicional de vacina de hepatite B (grupo 3). Após um período mediano de 10 anos, 54/70 (77,1%) destes indivíduos mantinham anti- HBs >= 10 mUI/mL. CONCLUSÕES: A avaliação dos marcadores sorológicos para VHB em PVHA vacinadas previamente para hepatite B evidenciou: alta persistência de anti-HBs após um período de 10 a 11 anos; doses adicionais de vacina foram capazes de induzir resposta humoral em indivíduos inicialmente não respondedores; não foram detectados marcadores sorológicos de infecção (HbsAg ou Anti-HBc) após 11 anos da vacinação inicial / BACKGROUND: Hepatitis B Virus (HBV) infection is responsible for great part of chronic hepatic diseases worldwide. In people living with HIV (PLHIV), HBV infection has more risk of progressing to cirrhosis and hepatocarcinoma. Hepatitis B vaccine is important in the prevention of a potentially severe disease, particularly in PLHIV, since both viruses have the same routes of transmission and co-infection has greater morbidity. Immunocompetent individuals have a good humoral response after the first hepatitis B vaccine series and no recommendation is made regarding booster doses. PLHIV may have a poor hepatitis B vaccine response, when compared to immunocompetent and the duration of immunity in these patients is unknown. OBJECTIVES: General: Evaluate serological markers of infection and protection from HBV in PLHIV previously vaccinated for hepatitis B. Specific: Evaluate anti-HBs persistence in PLHIV previously vaccinated for HBV who responded to a primary vaccine series; evaluate response to revaccination for hepatitis B in patients who did not respond to first vaccine series; investigate serological markers of infection from HBV in PLHIV previously vaccinated for hepatitis B. METHODS: Observational retrospective study of a PLHIV cohort primarily vaccinated between 2001 and 2002 for hepatitis B. Serological markers of infection and protection from HBV were investigated in those patients who were still attending the HIV/AIDS Patient Care Extension Service at the Clinical Division of Infectious and Parasitic Diseases attached to Hospital das Clínicas at Faculdade de Medicina at Universidade de São Paulo between 2012 and 2014. RESULTS: A cohort of 121 PLHIV was analyzed for seroconversion and persistence of anti-HBs. The majority were female (54.5%) and mean age 50.1 years. From these patients, 58 (group 1) were initially responders to the first vaccine series (anti- HBs >=10 mIU/mL) and 63 (group 2) were non- responders. After a median period of 11 years, none of the patients had serologic evidence of HBV infection and 41/58 (70.7%) of the initially responders had maintained anti-HBs >=10 mIU/mL. Greater CD4+ cell counts and anti- HBs>= 100mIU/mL at the time of first vaccine series were associated with persistence of anti-HBs. During evaluation period, 35/63 (55.6%) of the initially non-responders (group 2) successfully seroconverted (anti-HBs >=10 mIU/mL) in response to one or more booster doses. Booster doses may be effective in PLHIV. Number of booster doses were associated to seroconversion. Seventy of the 121 patients did not receive any further booster doses of hepatitis B vaccine from the time of their seroconversion (anti-HBs >=10 mIU/mL) (group 3). After 10 years of the seroconversion, 54/70 (77,1%) of these individuals has maintained anti- HBs >= 10 mIU/mL. CONCLUSIONS: Evaluation of serological markers for HBV in PLHIV previously vaccinated for hepatitis B showed: strong persistence of anti-HBs after a period of 10 to 11 years; additional vaccine doses elicited humoral response in initially non-responders; there was no serologic evidence of HBV infection (HbsAg ou Anti-HBc) about 11 years after initial vaccination

by Lau Tze Chin, Gene. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1991. / Includes bibliographical references (Leaves 170-195). / Abstract --- p.1 / Acknowledgement --- p.3 / List of tables --- p.4 / List of figures --- p.6 / List of abbreviations --- p.7 / Chapter Chapter One - --- Introduction --- p.9 / Chapter 1.1. --- Historical Aspects --- p.9 / Chapter 1.2. --- Classification of hepatitis B virus --- p.12 / Chapter 1.2.1. --- Hepadnaviruses --- p.12 / Chapter 1.2.2. --- Comparative properties of hepadnaviruses --- p.13 / Chapter 1.2.2.1. --- Physical properties --- p.13 / Chapter 1.2.2.2. --- Genetic relatedness --- p.15 / Chapter 1.2.2.3. --- Pathogenesis --- p.16 / Chapter 1.3. --- Structural and morphological properties of HBV --- p.17 / Chapter 1.4. --- Molecular biology of HBV --- p.20 / Chapter 1.4.1. --- Molecular structure of HBV --- p.20 / Chapter 1.4.1.1. --- Biochemistry of the virion envelope --- p.20 / Chapter 1.4.1.2. --- The nucleocapsid --- p.21 / Chapter 1.4.1.3. --- Structural features of HBV genome --- p.23 / Chapter 1.4.2. --- Genetic organization of HBV --- p.24 / Chapter 1.4.3. --- Infection cycle of HBV --- p.29 / Chapter 1.4.3.1. --- Viral attachment and internalization --- p.29 / Chapter 1.4.3.2. --- Replication of HBV --- p.30 / Chapter 1.4.3.3. --- Gene expression and regulation --- p.31 / Chapter 1.4.3.4. --- Host-virus DNA interaction --- p.33 / Chapter 1.5. --- Epidemiology and transmission of HBV --- p.34 / Chapter 1.5.1. --- World wide prevalence --- p.35 / Chapter 1.5.1.1. --- HBsAg prevalence --- p.35 / Chapter 1.5.1.2. --- Cumulative rate of HBV infection --- p.35 / Chapter 1.5.1.3. --- Age specific pattern of HBV infection --- p.36 / Chapter 1.5.2. --- Epidemiological pattern of HBV in Hong Kong --- p.37 / Chapter 1.5.3. --- Mode of transmission --- p.38 / Chapter 1.6. --- Clinical outcomes of HBV infection --- p.38 / Chapter 1.6.1. --- Acute infection --- p.41 / Chapter 1.6.2. --- Chronic infection --- p.42 / Chapter 1.6.3. --- Primary hepatocellular carcinoma --- p.43 / Chapter 1.7. --- Laboratory diagnosis of hepatitis B --- p.44 / Chapter 1.7.1. --- The HBV markers --- p.47 / Chapter 1.7.1.1. --- HBsAg and anti-HBs --- p.47 / Chapter 1.7.1.2. --- HBcAg and Anti-HBc --- p.47 / Chapter 1.7.1.3. --- HBeAg and anti-HBe --- p.49 / Chapter 1.7.1.4. --- HBV-associated DM polymerase --- p.49 / Chapter 1.7.1.5. --- HBV-DNA --- p.49 / Chapter 1.7.2. --- Methodology in the detection of hepatitis B markers --- p.50 / Chapter 1.7.2.1. --- Direct detection of HBV and HBV antigens --- p.50 / Chapter 1.7.2.2. --- Serological detection of HBV markers --- p.51 / Chapter 1.7.2.3. --- HBV-associated DNA polymerase assay --- p.51 / Chapter 1.7.2.4. --- Molecular technique for the detection and quantitation of HBV-DNA --- p.52 / Chapter 1.8. --- Antiviral therapy in hepatitis B --- p.52 / Chapter 1.8.1. --- Therapeutic agents for treatment of HBV infection --- p.53 / Chapter 1.8.1.1. --- Steroids --- p.53 / Chapter 1.8.2.2. --- Nucleoside analogs --- p.54 / Chapter 1.8.1.3. --- Interferon --- p.55 / Chapter 1.8.2. --- Clinical trials of interferons --- p.55 / Chapter 1.9. --- Extrahepatic tissue tropism of HBV --- p.62 / Chapter 1.10. --- Objective and design of study --- p.65 / Chapter 1.10.1. --- Objectives of study --- p.65 / Chapter 1.10.2. --- Study design --- p.66 / Chapter 1.10.2.1. --- Cross-sectional study --- p.67 / Chapter 1.10.2.2. --- Longitudinal study --- p.67 / Chapter 2.1. --- Materials --- p.71 / Chapter 2.1.1. --- Patients recruitment and clinical materials --- p.71 / Chapter 2.1.1.1. --- Cross-sectional study --- p.71 / Chapter 2.1.1.2. --- Longitudinal study --- p.71 / Chapter 2.1.2. --- Bacteria] stock --- p.71 / Chapter 2.1.3. --- "Chemicals, equipments and consumables" --- p.72 / Chapter 2.1.4. --- Buffers and solutions --- p.72 / Chapter 2.1.4.1. --- Phosphate buffer saline (PBS) --- p.72 / Chapter 2.1.4.2. --- Leucocyte lysis buffer (X 5)(LLB) --- p.72 / Chapter 2.1.4.3. --- Buffer equilibrated phenol (BEP) --- p.76 / Chapter 2.1.4.4. --- Phenol-Chloroform mixture --- p.76 / Chapter 2.1.4.5. --- 3.0M sodium acetate (pH 5.2) --- p.76 / Chapter 2.1.4.6. --- Tris-EDTA buffer (pH 8.0) (TE) --- p.76 / Chapter 2.1.4.7. --- Stock salmom sperm DNA solution --- p.77 / Chapter 2.1.4.8. --- Tracking dye --- p.77 / Chapter 2.1.4.9. --- Tris-borate electrophoresis buffer (TBE) --- p.77 / Chapter 2.1.4.10. --- Luria-Bertani Broth (LB) --- p.77 / Chapter 2.1.4.11. --- Solution ] --- p.78 / Chapter 2.1.4.12. --- Solution ]] --- p.78 / Chapter 2.1.4.13. --- Potassium acetate buffer (pH 5.4) --- p.78 / Chapter 2.1.4.14. --- Column elution buffer (CEB) --- p.78 / Chapter 2.1.4.15. --- NPMEB solution --- p.79 / Chapter 2.1.4.16. --- Neutralizing solution --- p.79 / Chapter 2.1.4.17. --- Standard saline citrate (SSC) --- p.79 / Chapter 2.1.4.18. --- Denhardt solution --- p.79 / Chapter 2.1.4.19. --- Prehybridization solution (PS) --- p.80 / Chapter 2.1.4.20. --- NETFAP Solution --- p.80 / Chapter 2.1.4.21. --- Heparin solution --- p.81 / Chapter 2.1.4.22. --- Hybridization mix for oligo-nucleotide probe --- p.81 / Chapter 2.1.4.23. --- NEPS solution (pH 7.0) --- p.81 / Chapter 2.1.4.24. --- Restriction endonuclease and buffer --- p.82 / Chapter 2.2. --- Methods --- p.82 / Chapter 2.2.1. --- Sample preparations --- p.82 / Chapter 2.2.1.1. --- Isolation of plasma and peripheral blood leucocytes (PBL) --- p.82 / Chapter 2.2.1.2. --- Extraction of DNA from Peripheral blood leucocytes --- p.83 / Chapter 2.2.1.3. --- Quantitation of Peripheral blood leucocyte DNA --- p.83 / Chapter 2.2.2. --- Preparation of radio-labelled HBV-DNA probe --- p.84 / Chapter 2.2.2.1. --- Plating and selection of bacterial stock --- p.84 / Chapter 2.2.2.2. --- Growth of E. coli HB101 and amplification of pAM6 --- p.84 / Chapter 2.2.2.3. --- Harvesting of E. coli and extraction of plasmid pAM6 --- p.84 / Chapter 2.2.2.4. --- Purification of plasmid pAM6 --- p.86 / Chapter 2.2.2.5. --- Large scale isolation and purification of HBV genome from plasmid pAM6 --- p.86 / Chapter 2.2.2.6. --- Radio-labelling of HBV-DNA --- p.88 / Chapter 2.2.2.6.1. --- Nick-translation of total HBV-DNA genome --- p.88 / Chapter 2.2.2.6.2. --- Multi-primer labelling of total HBV- DNA genome --- p.88 / Chapter 2.2.2.6.3. --- End-labeling of 21-base HBV oligo- nucleotide --- p.88 / Chapter 2.2.2.6.4. --- Determination of labelling efficiency --- p.89 / Chapter 2.2.2.7. --- Purification of labelled HBV-DNA probe --- p.90 / Chapter 2.2.2.7.1. --- Total genomic HBV-DNA probe (pAM6 probe) --- p.90 / Chapter 2.2.2.7.2. --- Oligo-nucleotide HBV-DNA probe (oligo probe) --- p.90 / Chapter 2.2.3. --- Hybridization study of clinical samples --- p.91 / Chapter 2.2.3.1. --- Solution hybridization of sera samples --- p.91 / Chapter 2.2.3.2. --- Spot hybridization of sera samples --- p.91 / Chapter 2.2.3.2.1. --- "Pre-hybridization treatment of sera samples (adapted from Lin et al.,1987)" --- p.91 / Chapter 2.2.3.2.2. --- Pre-hybridization and hybridization of the membrane --- p.92 / Chapter 2.2.3.2.3. --- Washing of membrane --- p.92 / Chapter 2.2.3.2.4. --- Final treatment and autoradiography: --- p.92 / Chapter 2.2.3.3. --- Quantitation of HBV-DNA in the sera samples: --- p.93 / Chapter 2.2.4. --- Assay for serological Hepatitis B marker --- p.93 / Chapter Chapter Three - --- Results --- p.93 / Chapter 3.1. --- Preparation of HBV-DNA probes --- p.95 / Chapter 3.2. --- Radiolabelling of HBV-DNA --- p.95 / Chapter 3.3. --- Hybridization methodology --- p.98 / Chapter 3.4. --- Comparison of the performance of HBV-DNA probes --- p.100 / Chapter 3.4.1. --- Quantitation of serum HBV-DNA --- p.100 / Chapter 3.4.2. --- Comparative hybridization performance of different HBV-DNA probes --- p.105 / Chapter 3.5. --- Clinical application of HBV-DNA probe:Detection of HBV-DNAin serum and peripheral blood leucocytes (PBL) --- p.109 / Chapter 3.5.1. --- Cross-sectional study --- p.112 / Chapter 3.5.1.1. --- Frequency of HBV-DNA detection in relation to different clinical manifestations --- p.112 / Chapter 3.5.1.2. --- Frequency of HBV-DNA detection in relation to the serological status --- p.114 / Chapter 3.5.1.3. --- Distribution of serum and PBL HBV-DNA level in chronic hepatitis B patients in relation to the different HBV-related manifestations --- p.119 / Chapter 3.5.2. --- Longitudinal study of patients with chronic hepatitis B under interferon therapy with prednisolone pretreatment --- p.123 / Chapter 3.5.2.1. --- Features of patients under study --- p.123 / Chapter 3.5.2.2. --- Correlation between the occurrence of HBV- DNA and HBeAg in serum --- p.123 / Chapter 3.5.2.3. --- Outcome of clinical trial: --- p.126 / Chapter 3.5.2.3.1. --- Number of patients responding to therapy: --- p.126 / Chapter 3.5.2.3.2. --- Variation in serum HBV markers during the course of study --- p.128 / Chapter 3.5.2.3.3. --- Change of HBV-DNA statusin peripheral blood leucocytes --- p.134 / Chapter Chapter Four - --- Dicussion --- p.140 / Chapter 4.1. --- Preparation of HBV-DNA hybridization probes --- p.140 / Chapter 4.1.1. --- Source of HBV-DNA --- p.140 / Chapter 4.1.2. --- Raidolabelling of HBV-DNA --- p.141 / Chapter 4.2. --- Hybridization methodology --- p.141 / Chapter 4.2.1. --- Optimization of hybridization conditions --- p.141 / Chapter 4.2.2. --- Comparison of the performance among different HBV- DNA probes --- p.144 / Chapter 4.3. --- Detection of HBV-DNA in clinical serum samples --- p.148 / Chapter 4.3.1. --- Crossectional study of patients with various categories of HBV related diseases --- p.148 / Chapter 4.3.1.1. --- HBV-DNA detection in serum --- p.148 / Chapter 4.3.1.2. --- Detection of HBV-DNA in peripheral blood mononuclear cells --- p.153 / Chapter 4.3.2. --- Longitudinal studies of patients undergoing antiviral therapy --- p.159 / Chapter 4.3.2.1. --- Serum HBV-DNA and HBeAg --- p.159 / Chapter 4.3.2.2. --- HBV-DNA in peripheral blood leucocytes --- p.163 / Conclusion --- p.166 / Future perspectives --- p.168 / References --- p.170

Concerns about increased stratospheric ozone depletion increasing ambient levels of ultraviolet-B radiation (UV-B), and the fact that some ecosystems are naturally exposed to high levels, has resulted in an increased interest in the effects of UV-B on plant communities. Despite this, there has been a paucity of studies into its effects on plant competition. Artificial plant communities consisting of Lolium perenne and Lotus corniculatus and a sub-montane community consisting of Agrostis tenuis, Festuca ovina and Galium saxatile (also including different nitrogen levels) were created using the response surface approach. The long-term effects of UV-B were also studied on a natural sub-Arctic community in Abisko, Sweden. In addition, all plant samples were analysed by Fourier-Transform Infrared Spectroscopy (FT-IR) to obtain a ‘metabolic fingerprint’ which was used to detect chemical differences to the whole biochemical complement of the sample. The results showed that enhanced UV-B altered the competitive interaction of Lolium perenne and Lotus corniculatus in favour of Lolium perenne although ambient levels of UV-B did not elicit an effect in the sub-montane community. Only one dwarf shrub species in the sub-Arctic experiment, Vaccinium myrtillus, was negatively affected by UV-B. In most cases, elevated UV-B elicited a change in the metabolic fingerprint in the samples and in some cases an alteration in competitive stress altered the metabolome. This suggests that FT-IR can be used as a screening tool to detect for both abiotic stress and competitive biochemical alterations. In addition, this thesis proposes that the facilitative effect between the grass-legume mixture of Lolium perenne and Lotus corniculatus is not related to nitrogen fixation in the early stages of competition which has traditionally been believed.

In this thesis I defend the principle of global egalitarianism. According to this idea most of the existing detrimental inequalities in this world are morally objectionable. As detrimental inequalities I understand those that are not to the benefit of the worst off people and that can be non-wastefully removed. To begin with, I consider various justifications of the idea that only those detrimental inequalities that occur within one and the same state are morally objectionable. I identify Thomas Nagel’s approach as the most promising defence of this traditional position. However, I also show that Nagel’s argument does not even justify the elimination of detrimental inequalities (that is to say: egalitarian duties of justice) within states. A discussion of the concept of political legitimacy rather shows that egalitarian justice is not a necessary condition of the justifiability of the exercise of coercive political power. I, then, consider other, more Rawlsian approaches to the question of detrimental inequalities. These views appear more plausible than Nagel’s position and argue that egalitarian duties also arise in certain international contexts. But also these more global theories of distributive justice suffer from shortcomings. Since they make the application of duties of justice dependent on the existence of social practices they cannot adequately account for the justified interests of non-participants that are affected by these practices. The counter-intuitive implications of practice-dependent theories lead me to investigate the plausibility of a theory that does not limit justice to existing practices and that argues for the inherent value of equality. This theory is global egalitarianism. I defend global egalitarianism by debilitating three objections that opponents of this idea frequently (but often not clearly) present in the relevant literature. Finally I also address two particular objections to the idea that global egalitarian duties are institutionalizable with the help of coercive global authorities.