Taiwan is high prevalence area in viral hepatitis. The prevalence rate is 17.3% in HBV but 4.4% in HCV. Taiwan government has started HBV vaccine policy since 1984. They hope to terminate the vertical infection and get good effectiveness. Now, HCV prevention is a important program for Taiwan. This survey wants to understand the donors¡¦ knowledge about HBV and HCV who live in Kaohsiung and Pingtung area. There are 321 available questionnaires. Male are 117(50.6%) and female are 114(49.4%). The result shows that cognition about HBV is better than HCV. The question ¡V ¡§HBV could be infected if dinner with someone who is HBV carrier.¡¨ is given false almost 47.2% but 47.6% in HCV. The donors who believe it will be infected HBV by donate blood are over 50% and same as HCV. The results show that donors have some mistake concept in transmission path, particularly in the safety about donate blood.

Background: A third of HIV-infected patients are co-infected with HCV in the developed world, and more of co-infected patients than ever before are dying because of liver related diseases today. Drug-related hepatotoxicity is a growing concern among human immunodeficiency virus (HIV) / hepatitis C virus (HCV) co-infected population. Nucleotide analogues containing HIV antiretroviral therapy, namely highly active anti-retroviral therapy (HAART), can induce mitochondrial toxicity. However, little is known about the effect of nucleotide analogues on the liver at the cellular and molecular level, and how it may affect treatment. Objective: To investigate whether liver tissue from HIV/HCV co-infected individuals will show greater liver mitochondrial toxicity if currently receiving antiviral HIV medication, compared to those who are not taking it. Methods: Liver biopsies were collected from 23 HIV/HCV co-infected males. Fourteen patients were on stable HAART (ON-HAART) and 9 were OFF-HAART, including 4 who stopped HAART >6 months prior and 5 who were HAART-nave. Liver mitochondrial toxicity was assessed by transmission electron microscopy-based quantitative stereological analyses of hepatocyte and mitochondrial morphometry, as well as by mitochondrial DNA (mtDNA) and mtRNA (COX1/(ß-actin) real-time-PCR quantification. Results: Hepatocytes tended to be larger in the ON-HAART group than in the OFF-HAART group (p=0.05), but they both showed similar mitochondrial volume fraction of the cell and mitochondrial crista density. Liver mtDNA and mtRNA levels were not significantly differentbetween ON-HAART and OFF-HAART. Hepatocyte lipid accumulation was significantly higher in HCV genotype 3 compared to genotype 1 infection ()=0.002), but was not associated with HAART status. Conclusions: We found no evidence or trend of increased mitochondrial toxicity in HIV/HCV co-infected individuals currently on HAART compared to those who are not. This finding could be relevant to the decision-making process with respect to initiating HCV therapy in this population.

NK cells control viral replication through cytotoxicity and IFNγ production. These functions were assessed in chronic HCV infected patients undergoing treatment. Aboriginals have genetic polymorphisms that may enhance NK cell function suggesting more effective clearance of chronic HCV than Caucasians. NK cell function was similar at baseline between ethnicities. At 3 months of treatment, Caucasian had higher NK killing potential compared to Aboriginal patients. This had no effect on treatment outcomes. NK cell cytotoxicity negatively correlated with viral loads while NK IFNγ production, particularly within the CD56bright subset, positively correlated with viral load suggesting that viral loads control NK cells function through an unknown mechanism. NK cell killing reflect fibrosis, but not liver damage measured by liver enzymes. IFNγ production,by NK cells does not reflect fibrosis nor liver enzymes levels. Lastly, NK cell function does not associate with therapeutic outcomes of chronic HCV infection suggesting that they do not directly play a role in therapeutic clearance of HCV.

Viral hepatitis infection represents a significant epidemiological and economic burden on society. Following infection, some patients mount a blunted immune response to the virus, which ultimately can result in chronic infection. FGL2, a member of the fibrinogen-related protein superfamily, has been implicated in vitro in suppressing both innate and adaptive immune responses. In a murine model of acute viral hepatitis caused by Lymphocytic Choriomeningitis Virus strain WE, we demonstrate that FGL2 expressed by reticuloendothelial cells limits viral spread. When expressed by Treg cells FGL2 binds to FCγRIIB and prevents DC maturation and suppresses virus-specific T and B cell responses. We provide compelling evidence to suggest that hepatitis viruses utilize the FGL2-FCγRIIB pathway to evade immune detection. Inhibition of this pathway restores effective cellular and humoral antiviral immune responses towards hepatitis viruses.

Viral hepatitis infection represents a significant epidemiological and economic burden on society. Following infection, some patients mount a blunted immune response to the virus, which ultimately can result in chronic infection. FGL2, a member of the fibrinogen-related protein superfamily, has been implicated in vitro in suppressing both innate and adaptive immune responses. In a murine model of acute viral hepatitis caused by Lymphocytic Choriomeningitis Virus strain WE, we demonstrate that FGL2 expressed by reticuloendothelial cells limits viral spread. When expressed by Treg cells FGL2 binds to FCγRIIB and prevents DC maturation and suppresses virus-specific T and B cell responses. We provide compelling evidence to suggest that hepatitis viruses utilize the FGL2-FCγRIIB pathway to evade immune detection. Inhibition of this pathway restores effective cellular and humoral antiviral immune responses towards hepatitis viruses.

NK cells control viral replication through cytotoxicity and IFNγ production. These functions were assessed in chronic HCV infected patients undergoing treatment. Aboriginals have genetic polymorphisms that may enhance NK cell function suggesting more effective clearance of chronic HCV than Caucasians. NK cell function was similar at baseline between ethnicities. At 3 months of treatment, Caucasian had higher NK killing potential compared to Aboriginal patients. This had no effect on treatment outcomes. NK cell cytotoxicity negatively correlated with viral loads while NK IFNγ production, particularly within the CD56bright subset, positively correlated with viral load suggesting that viral loads control NK cells function through an unknown mechanism. NK cell killing reflect fibrosis, but not liver damage measured by liver enzymes. IFNγ production,by NK cells does not reflect fibrosis nor liver enzymes levels. Lastly, NK cell function does not associate with therapeutic outcomes of chronic HCV infection suggesting that they do not directly play a role in therapeutic clearance of HCV.

Hepatitis C virus (HCV), a positive-sense RNA virus that chronically infects between 2.7 and 3.9 million Americans, is highly mutational, making the HCV infection difficult to treat. Thus, it is of high interest to search for highly conserved therapeutic targets within the HCV genome. Two such sequences are located within the 5' untranslated region (UTR) of HCV, being complementary for the microRNA miR-122, a liver microRNA essential for the production of the infectious virus. The use of peptide nucleic acids (PNAs) as therapeutic agents has become a promising area of study in recent years. In this study, we characterized the interactions between miR-122 and the HCV 5'UTR and designed PNAs to disrupt these interactions and thus, inhibit RNA replication and translation. Our results show that the PNAs effectively disrupt the interactions involving miR-122 and the 5'UTR, thereby increasing the possibility of a new therapeutic option against HCV. / Bayer School of Natural and Environmental Sciences / Chemistry and Biochemistry / MS / Thesis

Background: A third of HIV-infected patients are co-infected with HCV in the developed world, and more of co-infected patients than ever before are dying because of liver related diseases today. Drug-related hepatotoxicity is a growing concern among human immunodeficiency virus (HIV) / hepatitis C virus (HCV) co-infected population. Nucleotide analogues containing HIV antiretroviral therapy, namely highly active anti-retroviral therapy (HAART), can induce mitochondrial toxicity. However, little is known about the effect of nucleotide analogues on the liver at the cellular and molecular level, and how it may affect treatment. Objective: To investigate whether liver tissue from HIV/HCV co-infected individuals will show greater liver mitochondrial toxicity if currently receiving antiviral HIV medication, compared to those who are not taking it. Methods: Liver biopsies were collected from 23 HIV/HCV co-infected males. Fourteen patients were on stable HAART (ON-HAART) and 9 were OFF-HAART, including 4 who stopped HAART >6 months prior and 5 who were HAART-nave. Liver mitochondrial toxicity was assessed by transmission electron microscopy-based quantitative stereological analyses of hepatocyte and mitochondrial morphometry, as well as by mitochondrial DNA (mtDNA) and mtRNA (COX1/(ß-actin) real-time-PCR quantification. Results: Hepatocytes tended to be larger in the ON-HAART group than in the OFF-HAART group (p=0.05), but they both showed similar mitochondrial volume fraction of the cell and mitochondrial crista density. Liver mtDNA and mtRNA levels were not significantly differentbetween ON-HAART and OFF-HAART. Hepatocyte lipid accumulation was significantly higher in HCV genotype 3 compared to genotype 1 infection ()=0.002), but was not associated with HAART status. Conclusions: We found no evidence or trend of increased mitochondrial toxicity in HIV/HCV co-infected individuals currently on HAART compared to those who are not. This finding could be relevant to the decision-making process with respect to initiating HCV therapy in this population.

Orientador: Rafael Plana Simões / Resumo: A Hepatite C é caracterizada por uma infecção crônica e progressiva causada pelo vírus da HCV. A proteína NS3 do vírus está envolvida no seu ciclo replicativo, sendo assim importante alvo de ação de antivirais denominados de ação direta. O inibidor da protease Boceprevir faz parte da primeira geração dessa classe de drogas. Porém, muitos pacientes apresentam resposta virológica não sustentada (RVNS) durante o tratamento devido à incapacidade de supressão da replicação viral. O objetivo desse trabalho foi avaliar a presença de mutações e polimorfismos genéticos na região codificadora da proteína NS3 do HCV em pacientes que apresentam indicação de uso do fármaco Boceprevir que estão associadas com à (RVNS). Amostras de 25 pacientes foram utilizadas para o sequenciamento da proteína NS3 do HCV e avaliadas de acordo com suas mutações através de análise filogenética e de modelagem molecular, mais especificamente, índice de reatividade e simulação de dinâmica molecular. A reconstrução filogenética e as análises de pressão de seleção mostraram que pacientes que apresentaram falha ou recidiva possuem vírus com algumas mutações em regiões na estrutura viral que são próximas aos sítios de ligação com o fármaco Boceprevir. As análises de índice de reatividade mostraram que a probabilidade da ligação covalente ocorrer entre ligante e receptor está associada com a probabilidade de desprotonação do aminoácido S139 da proteína NS3. Já os resultados de dinâmica molecular mostram que a estabi... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

Submitted by HELOÍSA DE CARVALHO SAMPAIO null (helocarvalhosampaio@yahoo.com.br) on 2017-04-20T18:23:22Z No. of bitstreams: 1 Heloisa_Carvalho_Sampaio CORREÇÕES Final.pdf: 2559595 bytes, checksum: 2ff544d86a396ad9c7a7fcf67d7af948 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-04-25T18:01:29Z (GMT) No. of bitstreams: 1 sampaio_hc_me_bot.pdf: 2559595 bytes, checksum: 2ff544d86a396ad9c7a7fcf67d7af948 (MD5) / Made available in DSpace on 2017-04-25T18:01:29Z (GMT). No. of bitstreams: 1 sampaio_hc_me_bot.pdf: 2559595 bytes, checksum: 2ff544d86a396ad9c7a7fcf67d7af948 (MD5) Previous issue date: 2017-02-23 / A Hepatite C é caracterizada por uma infecção crônica e progressiva causada pelo vírus da HCV. A proteína NS3 do vírus está envolvida no seu ciclo replicativo, sendo assim importante alvo de ação de antivirais denominados de ação direta. O inibidor da protease Boceprevir faz parte da primeira geração dessa classe de drogas. Porém, muitos pacientes apresentam resposta virológica não sustentada (RVNS) durante o tratamento devido à incapacidade de supressão da replicação viral. O objetivo desse trabalho foi avaliar a presença de mutações e polimorfismos genéticos na região codificadora da proteína NS3 do HCV em pacientes que apresentam indicação de uso do fármaco Boceprevir que estão associadas com à (RVNS). Amostras de 25 pacientes foram utilizadas para o sequenciamento da proteína NS3 do HCV e avaliadas de acordo com suas mutações através de análise filogenética e de modelagem molecular, mais especificamente, índice de reatividade e simulação de dinâmica molecular. A reconstrução filogenética e as análises de pressão de seleção mostraram que pacientes que apresentaram falha ou recidiva possuem vírus com algumas mutações em regiões na estrutura viral que são próximas aos sítios de ligação com o fármaco Boceprevir. As análises de índice de reatividade mostraram que a probabilidade da ligação covalente ocorrer entre ligante e receptor está associada com a probabilidade de desprotonação do aminoácido S139 da proteína NS3. Já os resultados de dinâmica molecular mostram que a estabilidade da interação fármaco e proteína não é alterada após mutações singulares, sugerindo que o mecanismo de resistência do HCV pode estar associado a uma combinação de mutações ou ainda a aspectos moleculares não analisados no presente trabalho, como mecanismos de acoplamento receptor e ligante e modos de vibração da proteína. / Hepatitis C is characterized by a chronic and progressive infection caused by the HCV virus. The NS3 protein of the virus is involved in its replicative cycle, thus being an important target for the action of so-called direct-acting antivirals. The protease inhibitor Boceprevir is part of the first generation of this class of drugs. However, many patients present unsupported virological response (UVR) during treatment because of inability to suppress viral replication. The objective of this work was to evaluate the presence of mutations and genetic polymorphisms in the coding of the VHC NS3 protein in patients with indication of use of the drug Boceprevir that are associated with unsustained virological response. Samples from 25 patients were used for the sequencing of the VHC NS3 protein and evaluated according to their mutations through phylogenetic analysis and molecular modeling, more specifically, reactivity index and molecular dynamics simulation. Phylogenetic reconstruction and selection pressure analyzes have shown that patients who have failed or relapse have viruses with some mutations in regions in the NS3 protein sequence that are close to the drug binding sites Boceprevir. Reactivity index analyzes have shown that the probability of covalent binding between ligand and receptor occurring is associated with the probability of deprotonation of the S139 amino acid of the NS3 protein. The molecular dynamics results show that the stability of the drug and protein interaction is not altered after single mutations, suggesting that the mechanism of resistance of HCV may be associated with a combination of mutations or molecular aspects not analyzed in the present work, as Receptor and ligand coupling mechanisms and modes of protein vibration.