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The leading causes of liver disease are Hepatitis C virus infection and chronic alcohol abuse. Alcohol accelerates liver disease in HCV but the mechanisms are poorly understood. The identification of molecular gene expression profiles on human liver tissue was performed using microarrays. Samples were obtained from alcoholic-cirrhotic, HCV-cirrhotic, HCV/alcohol-cirrhotic and control non-cirrhotic liver tissue. Probe set expression summaries were calculated using RMA. Probe set level linear models were fit where probe set expression was modeled by HCV status, alcohol status, and the interaction between HCV and Alcohol. HCV cirrhosis was associated with up-regulation of genes related to viral and immune response, apoptosis and inflammation. There were down-regulation of genes in the ubiquititin-proteasome system in alcoholic cirrhosis. The interaction of HCV and alcohol revealed negative interaction for genes involved in apoptosis and immune response. There was a negative estimate for genes involved in class II restricted antigen presentation.

Wong, Yee Man Melody. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 148-174). / Abstract also in Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / Abbreviations --- p.v / Review of the literature --- p.1 / Overview of liver fibrosis --- p.2 / Pathophysiology of liver fibrosis --- p.3 / Histological classification of liver fibrosis --- p.4 / Gold standard for fibrosis assessment - Liver biopsy --- p.6 / Biomarker in blood - non-invasive method for assessing diseases --- p.7 / Significance of non-invasive markers of liver fibrosis --- p.9 / Biomarkers of liver fibrosis --- p.10 / Direct markers --- p.10 / Indirect markers --- p.11 / Proteomics / Why proteomics? --- p.16 / Clinical values of proteomics in biomarker discovery --- p.17 / Challenges in proteomics --- p.18 / Current proteomics technologies in biomarker discovery --- p.21 / Gel based --- p.21 / Gel free approach - MS based --- p.23 / Quantitative proteomics --- p.29 / Application of proteomics to discovery of biomarkers for diagnosis of liver fibrosis --- p.33 / Rationale and Objectives of the Project --- p.35 / Chapter Section 1 --- Method development of magnetic beads-based proteomic profiling for quantitative proteomic profiling and micro- purification in parallel --- p.36 / Chapter 1.1 --- Introduction --- p.36 / Chapter 1.2 --- Materials and methods --- p.38 / Chapter 1.3 --- Results / Chapter 1.3.1 --- Serum proteome profiles obtained with different types of chromatographic magnetic beads --- p.46 / Chapter 1.3.2 --- Performance of PCS 4000 ProteinChip reader --- p.49 / Chapter 1.3.3 --- Reproducibility of magnetic beads-based serum proteomic profiling --- p.54 / Chapter 1.3.4 --- Gel electrophoresis of the eluted proteins --- p.58 / Chapter 1.3.5 --- Identification of the protein peaks --- p.58 / Chapter 1.4 --- Discussion --- p.60 / Chapter 1.5 --- Conclusion --- p.64 / Chapter Section 2 --- Development of a proteome-based fingerprinting model for detecting liver fibrosis in patients with chronic hepatitis B infection --- p.65 / Chapter 2.1 --- Introduction --- p.65 / Chapter 2.2 --- Materials and methods --- p.68 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Patients characteristics --- p.75 / Chapter 2.3.2 --- Correlation between biochemical/serological markers and the degrees of liver fibrosis --- p.81 / Chapter 2.3.3 --- Serum proteomic profiling by linear MALDI-TOF MS --- p.81 / Chapter 2.3.4 --- Correlation of proteomic features with Ishak score --- p.81 / Chapter 2.3.5 --- Correlation of significant proteomic features with serological markers --- p.89 / Chapter 2.3.6 --- Construction of diagnostic model in detecting liver fibrosis and cirrhosis --- p.91 / Chapter 2.3.7 --- Cross-validation of the diagnostic model using pre- treatment samples in detecting liver fibrosis and cirrhosis --- p.91 / Chapter 2.3.8 --- Independent validation of the diagnostic model using post-treatment samples in detecting liver fibrosis and cirrhosis --- p.95 / Chapter 2.3.9 --- Comparison against other non-invasive models in detecting liver fibrosis and cirrhosis --- p.98 / Chapter 2.4 --- Discussion --- p.103 / Chapter 2.5 --- Conclusion --- p.112 / Chapter Section 3 --- Identification of proteomic features to form diagnostic fingerprint for the detection of liver fibrosis in patients with chronic hepatitis B infection --- p.113 / Chapter 3.1 --- Introduction --- p.113 / Chapter 3.2 --- Materials and methods --- p.115 / Chapter 3.3 --- Results --- p.121 / Chapter 3.3.1 --- Protein identification of the protein marker in the diagnostic model --- p.121 / Chapter 3.3.2 --- Immunodepletion of apolipoprotein C-III --- p.125 / Chapter 3.3.3 --- Serum levels of apolipoprotins and their association with liver fibrosis --- p.127 / Chapter 3.4 --- Discussion --- p.130 / Chapter 3.5 --- Conclusion --- p.139 / General discussion --- p.141 / Reference --- p.148 / Original Data --- p.175

Leung, Kin Chung. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 148-164). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Diss. (sammanfattning) Göteborg : Göteborgs universtiet, 2008. / Härtill 4 uppsatser.

Background and Aim: Bacterial infections are a well described complication of cirrhosis and occur in 37% of hospitalized patients. Culture positive infections in addition to the presence of bacterial products and DNA lead to loss of liver function and decompensation in cirrhosis. The mechanisms and molecular pathways associated with Bacterial Translocation (BT) are unknown. The aims of this study were to determine: i. macrophage phenotype and molecular pathways associated with bacterial translocation ii. if intestinal macrophages in liver cirrhosis are capable of modulating intestinal permeability.iii. structural integrity of the epithelial barrier. Methods: Duodenal biopsies and serum samples were collected from 29 patients with decompensated cirrhosis, 15 patients with compensated and 19 controls. Duodenal macrophages were characterized by means of flow cytometry and IHC. Gene expression analysis was performed to determine molecular pathways involved in BT. Inflammatory cytokine determination was done in serum and culture supernatant by means of customized cytometric bead arrays. Results: Patients with decompensated cirrhosis demonstrated: increased frequency of CD33+/CD14+/TREM-1+ and iNOS+ macrophages in their duodenum, elevated mRNA levels of nitric oxide synthase 2 (NOS2), chemokine ligand 2 (CCL2), chemokine ligand 13 (CCL13) and interleukin 8 (IL8) and increased serum levels of interleukin 6 (IL6), IL8 and lipopolysaccharides (LPS). Additionally, patients with decompensated cirrhosis showed an increase in NO, IL6, IL8 and CCL2 levels in culture supernatant after short term duodenal biopsy culture. Although the epithelial barrier on EM seemed intact, significantly increased expression of the “pore” forming tight junction claudin 2 was observed. Conclusion: This study showed the presence of activated CD14+Trem- 1+iNOS+ intestinal macrophages and increased levels of NO, IL-6 and claudin-2 levels in the duodenum of patients with decompensated liver cirrhosis, suggesting that these factors enhance intestinal permeability to bacterial products. / Afrikaans: Inleiding: Bakteriele infeksie is ‘n beskryfde komplikasie van lewersirrose wat in 37% van gehospitaliseerde pasiente voorkom. Kultuur positiewe infeksies asook die teenwoordigheid van bakteriele produkte en DNA lei tot verlies van lewerfunksie en dekompensasie. Die molekulere meganismes wat verband hou met bakteriele translokasie is nog onbekend. Die doel van hierdie studie was om: i. Makrofaag fenotipe en molekulere meganismes geassosieerd met bakteriele translokasie te beskryf, ii. te bepaal of intestinale makrofage dermdeurlaatbaarheid beinvloed, asook iii. om die struktruele integriteit van die dermwand te bepaal. Methods: Serum en dunderm biopsies was verkry van 29 pasiente met gedekompenseerde lewer sirrose, 15 pasiente met gekompenseerde sirrose en 19 kontroles. Dunderm makrofage was gekarakteriseer met behulp van vloeisitometrie en immunohistochemie. Molekulere meganisms belangrik tydens bakteriele translokasie was bepaal met behulp van geneekspressie. Serum en selkultuur supernatant sitokien bepalings was met Bioplex assays gedoen. Resultate: Pasiente met gedekompenseerde sirrose demonstreer: ‘n verhoogde frekwensie van CD33+/CD14+/TREM-1+ en iNOS+ makrofage in hul dunderm, verhoogde mRNA vlakke van NOS2, CCL2, CCL13 en IL8 asook verhoogde serum vlakke van IL6, IL8, LPS. Addisioneel het pasiente met gedekompenseerde sirrose vehoogde supernatant vlakke van NO, IL6, IL8 and CCL2 na kort termyn dunderm biopsie kulture. Alhoewel elekronmikroskopie gewys het dat die dundermwand intak is, was daar statisties-beduidend verhoogde ekspressie van die “porie” vormende vasteaansluitings- proteien, claudin 2 sigbaar. Gevolgtrekking: Gesamentlik het die studie gewys dat geaktiveerde CD14+/Trem-1+/iNOS+ intestinale makrofage asook verhoogde vlakke van NO, IL-6 en claudin-2 teenwoordig is in die dunderm van pasiente met gedekompenseerde sirrose. Dit dui daarop dat diè faktore derm deurlaatbaarheid vir bakteriele produkte kan verhoog. / Dissertation (MSc)--University of Pretoria, 2011. / Immunology / MSc / Unrestricted

abstract / Medicine / Master / Master of Philosophy

Não consta resumo na publicação. / Abstract not available.

A doença hepática gordurosa não alcoólica (DHGNA) compreende um amplo espectro morfológico de doenças com potencial de progressão em pacientes sem história de etilismo. Pode evoluir para esteatohepatite não alcoólica (EHNA), fibrose, cirrose e carcinoma hepatocelular. Com intuito de investigar as diferenças genéticas entre tecido hepático normal e cirrose secundária a EHNA, utilizamos microarranjos de oligonucleotídeos (CodeLink Uniset Human Whole Genome Bioarray System GE Healthcare - Bio-Sciences, Chalfont St. Giles, UK) para caracterizar os perfis de expressão nas duas condições. Analisamos 3 amostras de cirrose secundária a EHNA e 3 amostras de fígado normal provenientes de doadores durante o transplante hepático. Para a análise dos dados utilizamos o programa GenesifterTM analysis (VizX Labs LLC, Seattle, WA, USA; http://www.genesifter.net) para identificar os genes diferencialmente expressos e também as vias biológicas moduladas de acordo com a ontologia gênica. Posteriormente, para avaliarmos a expressão imunohistoquímica utilizamos a técnica de microarranjo tecidual em amostras de fígado normal (n=12), cirrose secundária a EHNA (n=10) e cirroses de outras etiologias (n=37). Identificamos 244 genes significativamente alterados em pelo menos 2 vezes. Destes, 138 genes apresentavam-se com expressão aumentada e 106 genes com expressão diminuída na cirrose secundária a EHNA comparados ao fígado normal. Foram selecionadas 9 vias metabólicas significativamente desreguladas. Dentre estas vias identificadas na cirrose secundária a EHNA, selecionamos a via de sinalização do mTOR e seu efetor 4EBP-1 para a análise da expressão protéica. Houve aumento significativo na expressão de 4EBP-1 no fígado normal comparado às outras cirroses, assim como na cirrose secundária a EHNA versus outras cirroses. Quanto à forma fosforilada houve apenas diferença na expressão entre fígado normal e cirroses de outras etiologias. A expressão de mTOR mostrou aumento significativo entre cirroses de outras etiologias quando comparadas ao fígado normal e cirrose secundária a EHNA. A expressão de mTOR fosforilado foi maior na cirrose secundária a EHNA quando comparada às cirroses de outras etiologias e fígado normal. Estudos recentes têm sugerido o papel do mTOR na DHGNA e o presente estudo corrobora a participação desta via também na cirrose secundária a EHNA. A avaliação imunohistoquímica de 4EBP-1 e mTOR fosforilado pode ser útil clinicamente para o diagnóstico diferencial entre cirrose secundária a EHNA versus cirroses de outras etiologias, quando a etiologia é desconhecida / Non-alcoholic fatty liver disease (NAFLD) encompasses the whole spectrum of steatosis, non-alcoholic steatohepatitis (NASH), and NASH-related cirrhosis (NASH/Cir). NASH/Cir can progress to hepatocellular carcinoma and reoccur post transplantation. Although molecular advances have been made in this field, the patogenesis of NAFLD is not completely understood. In an effort to investigate genetic differences between normal liver and NASH/Cir, first we used cDNA microarray (CodeLink Uniset Human Whole Genome Bioarray System GE Healthcare - Bio-Sciences, Chalfont St. Giles, UK) in normal liver from donor liver wedge biopsies taken at transplantation (n=3) and confirmed NASH/Cir tissues (n=3) and GenesifterTM analysis (VizX Labs LLC, Seattle, WA, USA; http://www.genesifter.net) to identify differentially expressed genes and biological pathways according to gene ontology (GO). Second, tissue microarray was used to determine immunohistochemical expression in normal liver samples (n=12), NASH/Cir (n=10) and in cirrhosis of other etiologies (n=37). Analysis of microarray data resulted in 244 genes changed in at least 2-fold with statistically significant ratio: 138 and 106 genes were, respectively, up and downregulated in NASH/Cir in comparison to normal liver. GO analysis by GeneSifterTM software identified nine statistically significant pathways containing differentially expressed genes. Among the 9 pathways identified as significantly modulated in NASH/Cir, we selected mTOR pathway and its downstream effector for immunohistochemical analysis. There was a significant increase in the expression of 4EBP-1 in normal liver compared to the other cirrhosis, as well as to NASH/Cir versus other cirrhosis. The phosphorylated 4EBP-1 showed only difference in expression between normal liver and cirrhosis of other etiologies. The expression of mTOR showed a significant increase in other cirrhosis when compared with normal liver and NASH/Cir. The expression of phosphorylated mTOR was higher in NASH/Cir when compared to other cirrhosis and normal liver. Recent findings have suggested a role for the cellular nutrient sensor mTOR in NAFLD and the present study corroborates the participation of this pathway in NASH/Cir. 4EBP-1 and phospho-mTOR evaluation might be of clinical utility as differential diagnostic of NASH/Cir from other cirrhosis, without knowing etiology

Made available in DSpace on 2014-06-11T19:22:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-12Bitstream added on 2014-06-13T19:07:27Z : No. of bitstreams: 1 vulcano_dsb_me_botfm.pdf: 1165851 bytes, checksum: ae1702dfaee6063d32a01859daa38f20 (MD5) / A doença hepática crônica influencia direta e indiretamente o estado nutricional do paciente, sendo a desnutrição bastante prevalente nesta população. As causas da desnutrição são multifatoriais, relacionadas à alteração na ingestão, na absorção e no metabolismo dos macro e micronutrientes. O diagnóstico nutricional torna-se difícil, pois, não existe um método considerado padrão-ouro e os métodos disponíveis podem não ser fidedignos pela influência da própria doença no resultado. O objetivo do trabalho foi avaliar os indicadores do estado nutricional e a composição corporal dos pacientes portadores de cirrose hepática e relacioná-los com o sexo, a etiologia e a gravidade da doença hepática. Os indicadores nutricionais avaliados foram os antropométricos (Índice de Massa Corpórea específico para pacientes cirróticos, dobras cutâneas e circunferências), os funcionais (dinamometria e espessura do músculo adutor do polegar), a avaliação nutricional subjetiva global e os laboratoriais (albumina, hemoglobina, hematócrito e contagem total de linfócitos). Para a avaliação da composição corporal foi utilizada a bioimpedância elétrica unifrequencial (BIA) que mensurou a resistência (R), a reactância (Xc), o ângulo de fase (AF) e a massa celular corpórea (MCC). A gravidade da doença hepática foi estimada pelos critérios de Child-Pugh e pelo Meld (The model end-stage liver disease). Os indicadores do estado nutricional foram dicotomisados em: desnutrição e sem desnutrição ou com depleção e sem depleção. Os pacientes cirróticos de etiologia alcoólica apresentaram maior depleção em todos os parâmetros antropométricos, funcionais e na avaliação nutricional subjetiva global (ANSG), sendo estatisticamente significante para Circunferência do Braço (CB), Circunferência Muscular do Braço (CMB) e ANSG... / The chronic hepatic disease directly and indirectly influences on patient’s nutritional status and malnutrition is highly prevalent in this population. The causes of malnutrition are multifactorial, related to alterations on ingestion, absorption and metabolism of macro and micronutrients. The nutritional diagnosis is difficult, because there isn’t a gold standard method, and available methods are not reliable, once the disease itself interferes in the result. The objective of the work was to appraise nutritional status indicators and body composition of cirrhotic patients treated in the nutrition first aid posts of Faculdade de Medicina de Botucatu – UNESP, and relate them to gender, etiology and severity of the liver disease. The nutritional indicators appraised have been the anthropometrical (specific body mass index for cirrhotic patients, skinfolds and circumferences), the functional (dynamometry and thickness of the adductor pollicis muscles), the Subjective Global Assessment of Nutritional Status (SGA) and the laboratorial (albumin, hemoglobin, hematocrit, lymphocytes total count). On the appraisement of body composition, unifrequency bioelectrical impedance analysis (BIA) has measured resistance (R), reactance (Xc), phase angle (PA) and body cell mass (BCM). The severity of the liver disease has been estimated by the Child-Pugh score and by Meld (The Model end-stage liver disease). The nutritional status indicators have been dichotomized as following: malnutrition and without malnutrition, or with depletion and without depletion. Cirrhotic patients with alcoholic etiology have exhibited more depletion in all the anthropometrical and functional parameters and in the Subjective Global Assessment of Nutritional Status (SGA) and they have been statistically relevant for Arm Circumference (AC), Arm Muscular Circumference (AMC) and SGA... (Complete abstract click electronic access below)