Introdução: A Agamaglobulinemia, a Imunodeficiência Comum Variável (IDCV) e a Síndrome Hiper IgM (SHIGM) são imunodeficiência primárias predominantemente humorais que se beneficiam da reposição de imunoglobulina humana, com redução da morbimortalidade. No entanto, apesar da reposição adequada de imunoglobulina, complicações pulmonares podem ocorrer, influenciando o prognóstico destes pacientes. Objetivo: O objetivo do estudo foi descrever as alterações morfológicas e funcionais pulmonares em pacientes com hipogamaglobulinemia primária em tratamento com reposição de imunoglobulina humana. Métodos: Foram avaliados 30 pacientes (agamaglobulinemia n=14; IDCV n=9; SHIGM n=7) que receberam imunoglobulina e antibioticoterapia profilática regularmente. A avaliação utilizou dados dos prontuários sobre o início e a evolução da doença, bem como dados de espirometria e tomografia computadorizada de tórax. O escore de Bhalla foi aplicado à tomografia mais recente de cada um dos pacientes para correlacionar as alterações tomográficas pulmonares com os dados clínicos, resultados das espirometrias e ocorrência de processos infecciosos sino-pulmonares após o início da reposição de imunoglobulina. Para as análises estatísticas utilizou-se o programa SPSS 13.0, e valores de p<0,05 foram considerados estatisticamente significantes. As variáveis nominais foram comparadas através do teste de Fisher e as contínuas, através de testes não paramétricos (Mann-Whitney, Kruskal- Wallis e Wilcoxon). Para as correlações do escore de Bhalla com as demais variáveis foi utilizado o coeficiente de Spearman. Resultados: Houve diminuição na frequência de pneumonias (p<0,001) e aumento na frequência de sinusites (p<0,001) após o início da reposição de imunoglobulina. Distúrbios ventilatórios foram evidenciados em 14 dos 23 pacientes que puderam realizaram espirometria (7 obstrutivos, 5 restritivos e 2 inconclusivos). Pacientes com bronquiectasias ao diagnóstico e aqueles à primeira avaliação tomográfica apresentaram mediana de idade mais elevada ao diagnóstico (p=0,015 e p=0,001, respectivamente) e tempo mais prolongado entre o início dos sintomas e o diagnóstico que aqueles sem bronquiectasias (p=0,010 e p=0,001, respectivamente). Sete pacientes desenvolveram bronquiectasias durante o tratamento. Pacientes com bronquiectasias à avaliação final apresentaram maior frequência de sinusites antes do início da reposição de imunoglobulina que aqueles sem bronquiectasias (p=0,010). Houve correlação estatisticamente significante do escore de Bhalla com VEF1 pré e pós-broncodilatador (r= -0,778 e r= -0,837, respectivamente), CVF (r= -0,773), FEF25-75% (r= -0,571) e com a frequência de pneumonias após o início do tratamento (r= 0,561). Conclusões: O tratamento com reposição regular de imunoglobulina e antibioticoterapia profilática reduziu a frequência e gravidade das infecções pulmonares, porém não evitou a ocorrência de sinusites, o aparecimento de bronquiectasias nem de outras alterações morfológicas e funcionais pulmonares / Introduction: Agammaglobulinemia, Common Variable Immunodeficiency (CVID) and Hyper IgM Syndrome (HIGM) are predominantly antibody deficiencies that benefit from immunoglobulin replacement therapy, with reduction of their morbidity and mortality. Despite regular immunoglobulin replacement, pulmonary complications may occur in those patients, affecting their prognosis. Objective: The aim of this study was to describe tomographic and functional pulmonary abnormalities in patients with primary hypogammaglobulinemia receiving immunoglobulin replacement therapy. Methods: Thirty patients (agammaglobulinemia n=14, CVID n=9, HIGM n=7) receiving antimicrobial prophylaxis and regular immunoglobulin infusions were evaluated. Clinical records were reviewed to obtain data concerning the onset and evolution of the disease and the results of spirometry and computed tomography of the chest. Bhalla score was applied to the most recent tomography of each patient to correlate tomographic pulmonary abnormalities with clinical data, spirometry results and the occurrence of sinusal and pulmonary infections after the onset of the immunoglobulin replacement. Statistical analysis was performed using the software SPSS 13.0 and p values < 0.05 were interpreted as statistically significant. Nominal variables were tested using Fisher´s exact test and continuous variables were tested using non-parametric tests (Mann-Whitney, Kruskal-Wallis e Wilcoxon). Spearman coefficient was used to correlate Bhalla score with the other variables. Results: The frequency of pneumonias decreased (p<0.001) and the frequency of sinusitis increased (p<0.001) after the onset of immunoglobulin replacement. Pulmonary function was abnormal in 14 of 23 patients (7 obstructive, 5 restrictive, 2 inconclusive). Patients with bronchiectasis at diagnosis and those with bronchiectasis at the first tomographic evaluation presented higher median age at diagnosis (p=0.015 and p=0.001, respectively) and longer duration between the onset of symptoms and diagnosis than those without bronchiectasis (p=0.010 e p=0.001, respectively). Seven patients developed bronchiectasis during treatment. Patients with bronchiectasis at the last tomographic evaluation presented a higher frequency of sinusitis before therapy onset than those without bronchiectasis (p=0.001). Statistically significant correlation was found between Bhalla score and pre and post bronchodilator FEV1 (r= -0.778 and r= -0.837, respectively), FVC (r= -0.773) and FEF25-75% (r= -0.571) and between Bhalla score and the frequency of pneumonias after the onset of immunoglobulin replacement therapy (r=0.561). Conclusions: Immunoglobulin replacement therapy and antimicrobial prophylaxis reduced the frequency and severity of pulmonary infections but did not prevent the occurrence of sinusitis, the development of bronchiectasis or other morphological and functional pulmonary abnormalities

A dengue é uma doença que afeta milhões de pessoas e possui um número significativo de mortes. Não há nenhum tratamento vacinal legalizado para uso. As estratégias vacinais contra a dengue baseadas em proteínas não estruturais têm demonstrado serem mais seguras do que as baseadas em proteínas estruturais. A proteína não estrutural 5 (NS5) do vírus dengue é a proteína mais conservada entre os quatro sorotipos e desempenha um papel crucial na replicação viral. Neste estudo, foi gerada uma forma recombinante da NS5 expressa em E. coli com propriedades antigênicas preservados. As condições de cultura foram optimizadas, o que permitiu a expressão dessa proteína na forma solúvel. A imunização de camundongos Balb/c com a NS5 sozinha ou em combinação com um adjuvante (poli (I:C)) promoveu o aumento da sobrevida de camundongos após desafio letal com DENV2. A combinação da NS5 com poli (I:C) emulsionado em Montanide 720 levou a expansão de linfócitos T CD8+ específicos. Os resultados indicam que a proteína NS5 obtida preserva determinantes antigênicos da proteína nativa e pode ser uma ferramenta útil para estudos sobre a biologia do DENV, busca de drogas antivirais e desenvolvimento de vacinas. / Dengue fever is a disease affecting millions of people worldwide and causing a significant number of deaths. There are no effective treatments or vaccine approaches capable of preventing such infection. Anti-DENV vaccine strategies based on nonstructural proteins as antigens have been shown to be safer than those based on structural proteins. The DENV nonstructural protein 5 (NS5), plays a crucial role in viral replication. In this study, we generated a recombinant form of DENV2 NS5 expressed in E. coli in high amounts and with preserved antigenic properties with regard to the native protein. Culture conditions were optimized in order to allow expression of NS5 as a soluble protein. The immunization of Balb/c mice using this protein alone or in combination with poly (I:C) led to increased survival after intracranial challenge with the DENV2 JHA1 strain. The combination of the protein with poly (I:C) emulsified in Montanide 720 led to the activation of NS5-specific CD8+ T lymphocytes. Altogether, the results indicate that the recombinant NS5 protein preserves antigenic determinants of the native protein and may be a useful tool for studies dealing with the DENV\'s biology, search for anti-viral drugs and vaccine development.

As Imunodeficiências primárias (IDPs) incluem um grupo heterogêneo de doenças, que resultam de distúrbios do sistema imunológico, aumentando a susceptibilidade a infecções. Os avanços terapêuticos têm propiciado maior sobrevida destes pacientes, assim como necessidades nutricionais. O objetivo deste estudo foi comparar o estado nutricional nos diferentes grupos de IDP, avaliados através dos seguintes escores Z: peso, estatura e IMC para idade e da distribuição dos percentis de circunferência muscular do braço (CMB) e dobra cutânea triciptal (DCT) para idade. Também foram realizados exames laboratoriais relacionados à avaliação nutricional. Foram avaliados 72 pacientes da Unidade de Alergia e Imunologia do ICr- HCFMUSP com diagnóstico definitivo ou provável de IDP, com idade entre 3 e 19 anos, alocados em três grupos de IDP: humoral (n=44), fagócitos (n=12) e celular ou combinada (n=19). Os três grupos foram descritos através de suas idades, sexo, peso e estatura, e com base nestes dados, determinou-se o escore Z de estatura para idade e o escore Z de IMC para idade. Para cada um desses escores Z, foram calculados: a média, o desviopadrão e o intervalo de confiança 95%, sendo as médias obtidas comparadas por ANOVA e posteriormente o teste POST HOC de Tukey- Kramer. Para avaliação de ingestão alimentar foram realizados: o recordatório alimentar de 24 horas e o questionário de frequência alimentar. Observou-se que os grupos de IDP celular ou combinada e de fagócitos apresentaram médias de escore Z de estatura e IMC menores do que o grupo IDP predominantemente humoral. As médias de escore Z de peso para idade não apresentaram diferença estatisticamente significante, entretanto o grupo predominantemente humoral teve a maior média. No grupo de IDP predominantemente humoral não houve diferenças estatisticamente significantes entre os subgrupos. A CMB estava abaixo do percentil 15 em 25% dos pacientes do grupo IDP predominantemente humoral; 55,5% do grupo de IDP de fagócitos; 68,4% do grupo de IDP celular ou combinada. Em relação à DCT, estavam abaixo do percentil 15: 18,1% dos pacientes do grupo IDP predominantemente humoral; 11% do grupo de IDP de fagócitos; e 36,8% do grupo de IDP celular ou combinada. A ingestão protéica excedeu o recomendado em 77%, 100% e 84,2% dos pacientes nos grupos de IDP predominantemente humoral, IDP de fagócitos e IDP celular ou combinada, respectivamente. Os grupos de IDP predominantemente humoral, IDP de fagócitos e IDP celular ou combinada apresentaram elevadas concentrações de colesterol. Em conclusão, observou-se déficit do estado nutricional mais intenso nos pacientes com IDP celular ou combinada e de fagócitos. O aumento de colesterol e a redução da CMB podem sugerir um distúrbio metabólico relacionado à inflamação crônica dos pacientes imunodeficientes, com risco para doença cardiovascular no futuro. / Primary immunodeficiencies (PID) include a heterogeneous group of diseases with immunological system disturbance, increasing susceptibility to infections. The therapeutic advances have increased life span of these patients, as well as the nutritional needs. The aim of this study was to compare the nutritional status of patients from different immunodeficiency groups evaluated by the following Z scores means: weight for age, height for age, body mass index (BMI) for age and the percentiles distribution of upper arm muscle circumference (UAMC) and triceps skin fold thickness (TSF) for age. Laboratorial exams related to nutritional status were also carried out. Among patients from Allergy and Immunology Unit from ICr-HCFMUSP, 72 patients with definitive or probable diagnosis were evaluated, with age from 3 to 19 years allocated in 3 PID groups: humoral (n=44), phagocytes (n=12) and cellular or combined (n=19). These three groups were described through age, gender, weight and height. Through these data, were determined the weight for age, height for age, and BMI for age Z scores. There were calculated for each Z score: the mean, standard-deviation and confidence interval 95%, being the obtained means compared by ANOVA and after, the Tukey-Kramer post hoc test. There were evaluated for intake food: a 24-hour recall and a food frequency questionnaire. It was observed that the cellular or combined and phagocyte groups presented lower height for age and BMI for age Z score means than the humoral group. The weight for age Z score means didnt show significant statistical difference, however the humoral group had the highest mean. Among the subgroups from humoral group was not found significant statistical difference. The UAMC was below the 15th percentile mean in 25% of the humoral group, 55.5% from the phagocyte group and 68.4% from the cellular or combined group. About TSF, 18.1% of patients from humoral group, 11% from phagocyte group and 36.8% from the cellular or combined group were below 15th percentile. The protein intake exceeded the RDA in 77%, 100% and 84.2% in patients from humoral, phagocyte and cellular or combined groups, respectively. The humoral, phagocyte and cellular or combined groups presented high cholesterol levels. In conclusion, it was observed more severe nutritional impairment in cellular or combined and phagocyte PID than the humoral group. The high cholesterol level and the UAMC reduction may suggest a metabolic disturbance related to chronic inflammation in PID patients, with risk for cardiovascular chronic disease in the future.

by Chan Po-cheung. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 218-244). / Acknowledgement --- p.i / Abbreviations --- p.ii / Abstract --- p.iv / Table of content --- p.ix / Chapter Chapter 1. --- Biochemistry of Tumor Necrosis Factor --- p.1 / Chapter I. --- Introduction --- p.1 / Chapter 1.1 --- The discovery of tumor necrosis factor (TNF) --- p.1 / Chapter 1.2 --- TNF as an antitumor agent --- p.3 / Chapter 1.3 --- Production of TNF --- p.4 / Chapter 1.4 --- Structure of TNF --- p.5 / Chapter 1.5 --- TNF receptor --- p.6 / Chapter 1.6 --- Biological activities of TNF --- p.10 / Chapter 1.7 --- Anti-tumor activity of TNF --- p.14 / Chapter 1.7.1 --- In vitro studies --- p.14 / Chapter 1.7.1.1 --- Synergistic effect of other cytokines --- p.14 / Chapter 1.7.1.2 --- DNA damages --- p.15 / Chapter 1.7.1.3 --- Free radical generation --- p.15 / Chapter 1.7.1.4 --- Utilization of ATP --- p.16 / Chapter 1.7.1.5 --- Phospholipase A2 activation --- p.17 / Chapter 1.7.2 --- In vivo studies --- p.17 / Chapter 1.8 --- Clinical trials --- p.18 / Chapter Chapter 2. --- Materials and Methods --- p.20 / Chapter 2.1 --- Materials --- p.20 / Chapter 2.2 --- Solutions commonly used --- p.21 / Chapter 2.3 --- Methods and procedure --- p.23 / Chapter 2.3.1 --- Culture of L-929 cells --- p.23 / Chapter 2.3.2 --- Trypan Blue exclusion test --- p.23 / Chapter 2.3.3 --- Determination of viability of L-929 cells upon rhTNF treatment --- p.24 / Chapter 2.3.4 --- Determination of cellular cAMP level --- p.25 / Chapter 2.3.5 --- Determination of inositol phosphate turnover --- p.26 / Chapter 2.3.6 --- Use of fluorescence probe in the study of rhTNF mediated killing --- p.28 / Chapter 2.3.6.1 --- Determination of changes in internal pH of L-929 cells --- p.29 / Chapter 2.3.6.2 --- Determination of intracellular calcium level in L-929 cells --- p.30 / Chapter 2.3.6.3 --- Determination of membrane potential by fluorescence probes --- p.32 / Chapter 2.3.6.4 --- "Translocation of nucleolar protein, nucleophosmin (B23)in L-929 cells" --- p.32 / Chapter 2.3.6.5 --- Determination of calcium mobilization in L-929 cells by confocal microscopy --- p.34 / Chapter 2.3.6.6 --- Determination of protein kinase C and phospho-tyrosine kinase in L-929 cells --- p.34 / Chapter 2.3.7 --- Uptake of 45Ca2+ in L-929 cells --- p.35 / Chapter 2.3.8 --- Measurement of membrane potential by Patch-clamp assay --- p.36 / Chapter 2.3.9 --- Determination of tyrosine kinase activation by Western blotting --- p.36 / Chapter 2.3.10 --- Statistical analysis --- p.38 / Chapter Chanter 3. --- Effect of rhTNF treatment on nucleophosmin in L-929 cells --- p.39 / Chapter 3.1 --- Introduction --- p.39 / Chapter 3.2 --- Results --- p.43 / Chapter 3.2.1 --- Effect of TNF (in the presence or absence of actinomycin D) on the nucleophosmin translocation in L-929 cells --- p.43 / Chapter 3.2.2 --- Effect of actinomycin D on the TNF-mediated cytotoxicity on L-929 cells --- p.51 / Chapter 3.3 --- Discussion --- p.57 / Chapter Chapter 4. --- Changes in membrane potential and intracellular pH in rhTNF-mediated cytotoxicity in L-929 cells --- p.59 / Chapter 4.1 --- Introduction --- p.59 / Chapter 4.2 --- Results --- p.61 / Chapter 4.2.1 --- Effect of rhTNF on the membrane potential of L-929 cells determined by fluorescence method --- p.61 / Chapter 4.2.2 --- Effect of rhTNF on the membrane potential of L-929 cells determined by patch clamp technique --- p.64 / Chapter 4.2.3 --- "Effect of K+, Na+ and pH on the rhTNF-mediated cytotoxicity on L-929 cells" --- p.67 / Chapter 4.3 --- Discussion --- p.90 / Chapter Chapter 5. --- Effect of intracellular cAMP and cAMP-dependent protein kinase (PKA) on the rhTNF-mediated cytotoxicity on L-929 cells --- p.92 / Chapter 5.1 --- Introduction --- p.92 / Chapter 5.1.1 --- "GTP-binding protein (G protein), cAMP and protein kinase A" --- p.92 / Chapter 2.1.2 --- Role of cAMP as second messenger --- p.96 / Chapter 5.1.3 --- Bacterial toxin used for study of G-protein --- p.98 / Chapter 5.1.4 --- Effect of cAMP on rhTNF cytotoxicity --- p.99 / Chapter 5.1.5 --- Effect of cAMP-dependent protein kinase (PICA) on rhTNF cytotoxicity --- p.101 / Chapter 5.2 --- Results --- p.102 / Chapter 5.2.1 --- Cyclic-AMP (cAMP) level in rhTNF-treated L-929 cells --- p.102 / Chapter 5.2.2 --- Effect of intracellular cAMP level on rhTNF-mediated cytotoxicity on L-929 cells --- p.104 / Chapter 5.2.3 --- Effect of agonist and inhibitor of cAMP dependent protein kinase (protein kinase A) on rhTNF-mediated cytotoxicity on L-929 cells --- p.107 / Chapter 5.2.4 --- Effect of protein kinase A inhibitors on rhTNF-mediated cytotoxicity on L-929 cells --- p.111 / Chapter 5.3 --- Discussion --- p.118 / Chapter Chapter 6. --- "Role of intracellular free calcium, ions and calcium dependent response in rhTNF-mediated cytotoxicity on L-929 cells" --- p.121 / Chapter 6.1 --- Introduction --- p.121 / Chapter 6.1.1 --- Inositol triphosphate and intracellular free calcium --- p.121 / Chapter 6.1.2 --- Diacylglycerol --- p.131 / Chapter 6.1.3 --- Protein kinase C (PKC) --- p.131 / Chapter 6.1.4 --- Intracellular free calcium ions and protein kinase C --- p.134 / Chapter 6.1.5 --- Effect of intracellular free calcium ions and protein kinase C on TNF-mediated cytotoxicity --- p.135 / Chapter 6.1.6 --- Tyrosine kinase induced release of IP3 --- p.136 / Chapter 6.1.7 --- Calcium channels --- p.136 / Chapter 6.2 --- Result --- p.139 / Chapter 6.2.1 --- Effect of rhTNF on intracellular free [Ca2+] of L-929 cells --- p.141 / Chapter 6.2.2 --- Effect of calcium ion channel blockers on rhTNF-mediated cytotoxicity on L-929 cells --- p.148 / Chapter 6.2.3 --- Effect of protein kinase C (PKC) on rhTNF-mediated cytotoxicity on L-929 cells --- p.158 / Chapter 6.2.4 --- Immunofluorescence staining of PKC in rhTNF-treated L-929 cells --- p.162 / Chapter 6.2.5 --- Effect of calmodulin and calmodulin sensitive calcium ATPase on rhTNF-mediated cytotoxicity on L-929 cells --- p.165 / Chapter 6.2.6 --- Role of inositol triphosphate in rhTNF-mediated cytotoxicity on L-929 cells --- p.167 / Chapter 6.2.7 --- Role of tyrosine kinase activity in the rhTNF-mediated cytotoxicity on L-929 cells --- p.185 / Chapter 6.3 --- Discussion --- p.191 / Chapter Chapter 7. --- Effect of antioxidants on rhTNF-mediated cytotoxicity on L-929 cells --- p.195 / Chapter 7.1 --- Introduction: Oxygen free radicals as mediators of rhTNF-induced tumor cell necrosis --- p.195 / Chapter 7.2 --- Results --- p.199 / Chapter 7.3 --- Discussion --- p.203 / Chapter Chapter 8. --- General Discussion --- p.205 / Bibliography --- p.217

by Joyce Chui Kwan Ho. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 61-75). / Abstracts in English and Chinese. / Acknowledgments --- p.i / List of Abbreviations --- p.iii / Abstract / English --- p.1 / Chinese --- p.2 / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Tumor Formation --- p.3 / Chapter 1.2 --- Anti-tumor Pathways --- p.4 / Chapter 1.3 --- Aim of Project --- p.13 / Chapter Chapter 2 --- The In Vivo effect of Polysaccharopeptide / Chapter 2.1 --- Introduction --- p.15 / Chapter 2.2 --- Materials and Methods --- p.17 / Chapter 2.3 --- Results --- p.18 / Chapter 2.4 --- Discussion --- p.19 / Chapter Chapter 3 --- Cytotoxicity / Chapter 3.1 --- Introduction --- p.23 / Chapter 3.2 --- Materials and Methods --- p.26 / Chapter 3.3 --- Results --- p.28 / Chapter 3.4 --- Discussion --- p.28 / Chapter Chapter 4 --- Anti-angiogenic Effect / Chapter 4.1 --- Introduction --- p.30 / Chapter 4.2 --- Materials and Methods --- p.35 / Chapter 4.3 --- Results --- p.39 / Chapter 4.4 --- Discussion --- p.42 / Chapter Chapter 5 --- Immunomodulation / Chapter 5.1 --- Introduction --- p.45 / Chapter 5.2 --- Materials and Methods --- p.47 / Chapter 5.3 --- Results --- p.50 / Chapter 5.4 --- Discussion --- p.52 / Chapter Chapter 6 --- General Discussion --- p.57 / References --- p.61

by Cheung Yuen-Nei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 194-208). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Contents --- p.vi / List of Figures --- p.xii / List of Tables --- p.xv / Abbreviations --- p.xvi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1 --- Introduction --- p.2 / Chapter 1.1 --- Characteristic of Saponins --- p.3 / Chapter 1.1.1 --- Occurrence of Saponins --- p.3 / Chapter 1.1.2 --- General Properties of Saponins --- p.3 / Chapter 1.1.2.1 --- Emulsifying Agents --- p.3 / Chapter 1.2.2.2 --- Forming Complex with Cholesterol --- p.4 / Chapter 1.1.2.3 --- Hemolytic Property --- p.4 / Chapter 1.1.3 --- Structure of Saponins --- p.5 / Chapter 1.1.3.1 --- Categories of Saponins --- p.5 / Chapter 1.1.3.1.1 --- Triterpene Saponins --- p.5 / Chapter 1.1.3.1.2 --- Steroid Saponins --- p.5 / Chapter 1.1.3.2 --- Monodesmosidic and Bidesmosidic Saponins --- p.7 / Chapter 1.1.4 --- Biological and Pharmacological Properties of Saponins --- p.9 / Chapter 1.1.4.1 --- Anti-microbial Activity --- p.9 / Chapter 1.1.4.1.1 --- Anti-fungal Activities --- p.9 / Chapter 1.1.4.1.2 --- Anti-bacterial Activities --- p.10 / Chapter 1.1.4.1.3 --- Anti-viral Activities --- p.10 / Chapter 1.1.4.2 --- Insecticidal Activity --- p.10 / Chapter 1.1.4.3 --- Molluscicidal Activity --- p.10 / Chapter 1.1.4.4 --- Hypocholesterolemic Activity --- p.11 / Chapter 1.1.4.5 --- Anti-ulcer Activity --- p.11 / Chapter 1.1.4.6 --- Contraceptive Activity --- p.12 / Chapter 1.1.4.7 --- Immunomodulatory Activities --- p.12 / Chapter 1.1.4.7.1 --- Direct Immunostimulation --- p.12 / Chapter 1.1.4.7.2 --- Acting as Immuno-adjuvants --- p.13 / Chapter 1.1.4.8 --- Anti-tumor Activity --- p.14 / Chapter 1.1.4.8.1 --- Anti-carcinogenesis --- p.15 / Chapter 1.1.4.8.2 --- Suppression of Tumor Growth --- p.16 / Chapter 1.1.5 --- Anti-tumor Activity of Steroid Saponins --- p.18 / Chapter 1.1.5.1 --- Diosgenin Steroid Saponin --- p.18 / Chapter 1.1.5.2 --- Hong Kong Compounds --- p.18 / Chapter 1.1.5.3 --- Hong Kong18 --- p.21 / Chapter 1.2 --- Human Hepatocellular Carcinoma (HCC) --- p.24 / Chapter 1.2.1 --- The Incidence of Liver Cancer --- p.24 / Chapter 1.2.2 --- Classification of Liver Cancer --- p.24 / Chapter 1.2.3 --- Human Hepatocellular Carcinoma Cell Lines --- p.25 / Chapter 1.2.3.1 --- Human Hepatocellular Carcinoma Cell Line HepG2 --- p.25 / Chapter 1.2.3.2 --- Multidrug Resistant Human Hepatocellular Carcinoma Cell Line R-HepG2 --- p.27 / Chapter 1.2.3.2.1 --- Mechanisms of Multidrug Resistance --- p.28 / Chapter 1.2.3.2.2 --- Structure and Characteristics of P-glycoprotein --- p.29 / Chapter 1.2.3.2.3 --- Methods in Dealing with P-glycoprotein Over-expressed MDR Cells --- p.31 / Chapter 1.3 --- Objectives of the Project --- p.32 / Chapter 1.3.1 --- Study of the Anti-tumor Activities of Hong Kong 18 on Human Hepatocellular Carcinoma Cell Line HepG2 and Unravel the Underlying Mechanisms --- p.32 / Chapter 1.3.2 --- Study of the Anti-tumor Activities of Hong Kong 18on Multidrug Resistant Human Hepatocellular Carcinoma Cell Line R-HepG2 and Unravel the Underlying Mechanisms --- p.32 / Chapter Chapter 2 --- Materials and Methods --- p.33 / Chapter 2.1 --- Materials --- p.34 / Chapter 2.1.1 --- Cell Culture --- p.34 / Chapter 2.1.1.1 --- Cell Lines --- p.34 / Chapter 2.1.1.2 --- Culture Media --- p.35 / Chapter 2.1.2 --- Reagents and Buffers --- p.36 / Chapter 2.1.2.1 --- Phosphate Buffered Saline (PBS) --- p.36 / Chapter 2.1.2.2 --- Reagents and Buffers for DNA Fragmentation --- p.36 / Chapter 2.1.2.3 --- Reagents and Buffers for Western Analysis --- p.37 / Chapter 2.1.2.4 --- Reagents and Buffer for Caspases Activities --- p.39 / Chapter 2.1.2.5 --- Fluorescent Dyes used for Flow Cytometry --- p.39 / Chapter 2.1.3 --- Chemicals --- p.39 / Chapter 2.2 --- Methods --- p.46 / Chapter 2.2.1 --- MTT Assay --- p.46 / Chapter 2.2.2 --- Determination of Cell Viability --- p.46 / Chapter 2.2.3 --- Purification of Macrophages from balb/c Mice --- p.47 / Chapter 2.2.4 --- Hemolysis Assay --- p.47 / Chapter 2.2.5 --- In vivo Studies of the Toxicity of HK18 --- p.48 / Chapter 2.2.6 --- DNA Fragmentation Assay --- p.50 / Chapter 2.2.7 --- Detection of Apoptotic and Necrotic / Late Apoptotic Cells Death by Flow Cytometry with Annexin V-FITC / PI --- p.51 / Chapter 2.2.8 --- Detection of Mitochondrial Membrane Potential by JC-1 Fluorescent Dye --- p.52 / Chapter 2.2.9 --- Detection of Intracellular Ca Level by Flow Cytometry with Fluo-3 Fluorescent Dye --- p.52 / Chapter 2.2.10 --- Detection of Intracellular Hydrogen Peroxide Level by Flow Cytometry with DCF Fluorescence Dye --- p.53 / Chapter 2.2.11 --- Simultaneous Detection of Mitochondrial Membrane Potential and Intracellular Ca2+ or Mitochondrial Membrane Potential and Intracellular Hydrogen Peroxide --- p.54 / Chapter 2.2.12 --- Western Analysis --- p.55 / Chapter 2.2.12.1 --- Total Protein Extraction --- p.55 / Chapter 2.2.12.2 --- Extraction of Cytosolic Proteins --- p.59 / Chapter 2.2.13 --- Determination of Caspases Enzymatic Activity --- p.63 / Chapter 2.2.14 --- Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) --- p.67 / Chapter 2.2.14.1 --- RNA Extraction by TRIzol Reagent --- p.67 / Chapter 2.2.14.2 --- Reverse Transcription --- p.68 / Chapter 2.2.14.3 --- Polymerase Chain Reaction --- p.68 / Chapter 2.3 --- Statistic Analysis --- p.71 / Chapter Chapter 3 --- Cytotoxicity of HK18 --- p.72 / Chapter 3.1 --- Cytotoxicity of HK18 on HepG2 Cells --- p.73 / Chapter 3.1.1 --- Study of the Cytotoxic Activity of HK18 on HepG2 Cells by MTT Assay --- p.73 / Chapter 3.1.2 --- Study of the Cytotoxic Activity of HK18 on HepG2 Cells by Tryphan Blue Exclusion Assay --- p.76 / Chapter 3.2 --- Cytotoxicity of HK18 on R-HepG2 Cells --- p.78 / Chapter 3.2.1 --- Study of the Cytotoxic Activity of HK18 on R-HepG2 Cells by MTT Assay --- p.78 / Chapter 3.2.2 --- Study of the Cytotoxic Activity of HK18 on R-HepG2 Cells by Tryphan Blue Exclusion Assay --- p.81 / Chapter 3.3 --- Cytotoxicity of HK18 on Macrophages --- p.83 / Chapter 3.4 --- Hemolytic Activity of HK18 --- p.85 / Chapter 3.5 --- In vivo Study of the Toxicity of HK18 --- p.87 / Chapter Chapter 4 --- Mechanistic Study of HK18 on HepG2 Cells --- p.90 / Chapter 4.1 --- Hallmarks of Apoptosis Induced by HK18 on HepG2 Cells --- p.91 / Chapter 4.1.1 --- Induction of Phosphatidylserine Externalization by HK18 on HepG2 Cells --- p.91 / Chapter 4.1.2 --- Induction of DNA Fragmentation by HK18 of HepG2 Cells --- p.97 / Chapter 4.2 --- Study of the Underlying Mechanisms of HK18 Induced Apoptosis in HepG2 Cells --- p.99 / Chapter 4.2.1 --- The Role of Mitochondria in HK18 Induced Apoptosis of HepG2 Cells --- p.99 / Chapter 4.2.1.1 --- HK18 Induced Mitochondrial Membrane Depolarization in HepG2 Cells --- p.101 / Chapter 4.2.1.2 --- Addition of Bongkrekic Acid Reduced HK18 Cytotoxicity on HepG2 Cells --- p.105 / Chapter 4.2.1.3 --- Elevation of Intracellular Hydrogen Peroxide Level in HK18 Treated HepG2 Cells --- p.108 / Chapter 4.2.1.4 --- Elevation of Intracellular Ca2+ Level in HK18 Treated HepG2 Cells --- p.114 / Chapter 4.2.1.5 --- HK18 Induced Cytochrome c and AIF Released from Mitochondria of HepG2 Cells --- p.120 / Chapter 4.3 --- Downstream Biochemical Changes Induced by HK18 on HepG2 Cells --- p.123 / Chapter 4.3.1 --- Activation of Caspase 3 of HepG2 Cells by HK18 as Demonstrated by Western Blot --- p.123 / Chapter 4.3.2 --- Induction of Caspases Activities of HepG2 Cells by HK18 as Demonstrated by Enzymatic Activity Assays --- p.125 / Chapter 4.4 --- Down-regulation of Anti-apoptotic Bcl-2 Family Members of HepG2 Cells by HK18 --- p.129 / Chapter Chapter 5 --- Mechanistic Study of HK18 on R-HepG2 Cells --- p.133 / Chapter 5.1 --- Hallmarks of Apoptosis Induced by HK18 on R-HepG2 Cells --- p.134 / Chapter 5.1.1 --- Induction of Phosphatidylserine Externalization by HK18 on R-HepG2 Cells --- p.134 / Chapter 5.1.2 --- Induction of DNA Fragmentation by HK18 of R-HepG2 Cells --- p.137 / Chapter 5.2 --- Study of the Underlying Mechanisms of HK18 Induced Apoptosis in R-HepG2 Cells --- p.139 / Chapter 5.2.1 --- The Role of Mitochondria in HK18 Induced Apoptosis of R-HepG2 Cells --- p.139 / Chapter 5.2.1.1 --- HK18 Induced Mitochondrial Membrane Depolarization in R-HepG2 Cells --- p.139 / Chapter 5.2.1.2 --- Addition of Bongkrekic Acid Reduced HK18 Cytotoxicity on R-HepG2 Cells --- p.142 / Chapter 5.2.1.3 --- Elevation of Intracellular Hydrogen Peroxide Level in HK18 Treated R-HepG2 Cells --- p.144 / Chapter 5.2.1.4 --- Elevation of Intracellular Ca2+ Level in HK18 Treated R-HepG2 Cells --- p.146 / Chapter 5.3 --- Downstream Biochemical Changes Induced by HK18 on R-HepG2 Cells --- p.148 / Chapter 5.3.1 --- Activation of Caspase 3 of R-HepG2 Cells by HK18 as Demonstrated by Western Blot --- p.148 / Chapter 5.3.2 --- Induction of Caspases Activation of R-HepG2 Cells by HK18 as Demonstrated by Enzymatic Activity Assays --- p.150 / Chapter 5.4 --- Down-regulation of the Anti-apoptotic Bcl-2 Protein of R-HepG2 Cells by HK18 --- p.154 / Chapter 5.5 --- HK18 was Not a Substrate of P-glycoprotein Contents --- p.156 / Chapter Chapter 6 --- Discussion --- p.158 / Chapter 6.1 --- Cytotoxicity of HK18 --- p.159 / Chapter 6.1.1 --- HK18 was Cytotoxic to the Human Hepatocellular Carcinoma Cell Line HepG2 and Multidrug Resistant Human Hepatocellular Carcinoma Cell Line R-HepG2 --- p.159 / Chapter 6.1.2 --- Study of the Toxicity of HK18 --- p.160 / Chapter 6.2 --- Mechanistic Studies of the Cytotoxic Effects of HK18 on HepG2 Cells --- p.161 / Chapter 6.2.1 --- Apoptotic Cell Death Induction of HK18 on HepG2 Cells --- p.161 / Chapter 6.2.2 --- Studies of the Underlying Mechanisms of HK18 Induced Apoptosis of HepG2 Cells --- p.162 / Chapter 6.3 --- Mechanistic Studies of the Cytotoxic Effects of HK18 on R-HepG2 Cells --- p.181 / Chapter 6.3.1 --- Apoptotic Cell Death Induction of HK18 on R-HepG2 Cells --- p.181 / Chapter 6.3.2 --- Studies of the Underlying Mechanisms of HK18 Induced Apoptosis of HepG2 Cells --- p.181 / Chapter Chapter 7 --- Future Perspectives --- p.190 / Chapter Chapter 8 --- References --- p.193

by Ko Samuel. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 209-227). / Abstract also in Chinese. / Acknowledgements --- p.i / Abbreviations --- p.ii / Abstract --- p.vii / Abstract in Chinese --- p.x / List of Figures --- p.xiii / List of Tables --- p.xx / Publication --- p.xxi / Contents --- p.xxii / Chapter Chapter 1. --- General Introduction --- p.1 / Chapter 1.1 --- Tumor Necrosis Factor --- p.2 / Chapter 1.1.1 --- History of Tumor Necrosis Factor --- p.2 / Chapter 1.1.2 --- TNF Subtypes and Their Purification --- p.3 / Chapter 1.1.3 --- Release of TNF --- p.9 / Chapter 1.1.4 --- Biological Actions of TNF --- p.9 / Chapter 1.2 --- Tumor Necrosis Factor Receptor --- p.11 / Chapter 1.2.1 --- Purification of TNF Receptor --- p.11 / Chapter 1.2.2 --- Regulation of TNF Receptor --- p.14 / Chapter 1.2.3 --- "Functions of TNF Receptor 1,Receptor 2 and Soluble TNF Receptors" --- p.15 / Chapter 1.3 --- Possible Signal Transductions of Tumor Necrosis Factor-Alpha --- p.17 / Chapter 1.3.1 --- Activation of Phospholipase A2 Cascade --- p.18 / Chapter 1.3.2 --- Activation of Phospho lipase C Pathway --- p.19 / Chapter 1.3.3 --- Activation of Sphingomyelin Pathway --- p.20 / Chapter 1.3.4 --- Activation of Protein Kinase --- p.22 / Chapter 1.3.5 --- Activation of the Cascade of Death Domain --- p.23 / Chapter 1.4 --- Induction of Both Necrosis and Apoptosis by Tumor Necrosis Factor-Alpha --- p.25 / Chapter 1.4.1 --- Apoptosis Versus Necrosis --- p.25 / Chapter 1.4.2 --- TNF Can Induce Both Apoptosis and Necrosis --- p.27 / Chapter 1.5 --- Possible Mechanisms of Tumor Necrosis Factor-Alpha- Mediated Cytotoxicity --- p.27 / Chapter 1.5.1 --- Release of Reactive Oxygen Species --- p.28 / Chapter 1.5.2 --- Release of Intracellular Calcium --- p.31 / Chapter 1.5.3 --- Miscellaneous Mechanisms --- p.36 / Chapter 1.6 --- Objective of Studies --- p.37 / Chapter Chapter 2. --- Materials and Methods --- p.39 / Chapter 2.1 --- Materials --- p.40 / Chapter 2.1.1 --- Buffer --- p.40 / Chapter 2.1.2 --- Culture Media --- p.45 / Chapter 2.1.3 --- Chemicals --- p.46 / Chapter 2.1.4 --- Culture of Cells --- p.49 / Chapter 2.1.4.1 --- "Tumor Necrosis Factor-Alpha-Sensitive Cell Line, L929" --- p.49 / Chapter 2.1.4.2 --- "Tumor Necrosis Factor-Alpha-Resistant Cell Line, rL929, rL929-l IE and rL929-4F" --- p.50 / Chapter 2.2 --- Methods --- p.50 / Chapter 2.2.1 --- Agarose Gel Electrophoresis --- p.50 / Chapter 2.2.2 --- Cytotoxicity Assay --- p.52 / Chapter 2.2.3 --- Confocal Laser Scanning Microscopy --- p.53 / Chapter 2.2.4 --- Flow Cytometry --- p.57 / Chapter Chapter 3. --- Results --- p.65 / Chapter 3.1 --- Induction of Apoptosis in Tumor Necrosis Factor-Alpha- Treated L929 Cell --- p.66 / Chapter 3.1.1 --- Introduction --- p.66 / Chapter 3.1.2 --- TNF Induced DNA Fragmentation in L929 Cells --- p.67 / Chapter 3.2 --- Effect of Tumor Necrosis Factor-Alpha on Cell Cycle --- p.73 / Chapter 3.2.1 --- Introduction --- p.73 / Chapter 3.2.2 --- Effect of TNF on Cell Cycle --- p.75 / Chapter 3.3 --- Release of Reactive Oxygen Species in Tumor Necrosis Factor-Alpha Treatment --- p.79 / Chapter 3.3.1 --- Introduction --- p.79 / Chapter 3.3.2 --- Release of Reactive Oxygen Species in TNF- Treated L929 Cells is Time Dependent --- p.81 / Chapter 3.3.3 --- Effect of Antioxidants on TNF-Mediated Cytotoxicity --- p.93 / Chapter 3.3.4 --- Effect of Mitochondrial Inhibitors on TNF-Mediated Cytotoxicity --- p.96 / Chapter 3.4 --- The Role of Calcium in Tumor Necrosis Factor-Alpha Treatment --- p.112 / Chapter 3.4.1 --- Introduction --- p.112 / Chapter 3.4.2 --- Release of Intracellular Calcium in TNF-Treated L929 Cells --- p.113 / Chapter 3.4.3 --- Effect of Calcium-Inducing Agents on TNF-Treated L929Cells --- p.127 / Chapter 3.5 --- Relationship between Reactive Oxygen Species and Calcium in Tumor Necrosis Factor-Alpha-Mediated Cytotoxicity --- p.133 / Chapter 3.5.1 --- Introduction --- p.133 / Chapter 3.5.2 --- Effect of Intracellular Calcium Chelator on TNF- Mediated ROS Release and Cytotoxicity --- p.133 / Chapter 3.5.3 --- Effect of Mitochondrial Calcium on TNF-Mediated ROS Release and Cytotoxicity --- p.147 / Chapter 3.6 --- Effect of Tumor Necrosis Factor-Alpha on pH --- p.162 / Chapter 3.6.1 --- Introduction --- p.162 / Chapter 3.6.2 --- Effect of TNF on pH --- p.162 / Chapter 3.7 --- Effect of Tumor Necrosis Factor-Alpha on Mitochondrial Membrane Potential --- p.165 / Chapter 3.7.1 --- Introduction --- p.165 / Chapter 3.7.2 --- Effect of TNF and Some Drugs on Mitochondrial Membrane Potential --- p.165 / Chapter 3.8 --- "Comparison of Effects of Tumor Necrosis Factor-Alpha on Susceptible Cell Line, L929 and Resistant Cell Line, rL929, rL929-11E and rL929-4F" --- p.169 / Chapter 3.8.1 --- Introduction --- p.169 / Chapter 3.8.2 --- Effect of TNF on the Cytotoxicity of Resistant Cell Lines --- p.170 / Chapter 3.8.3 --- Effect of TNF on the Release of ROS in Resistant Cell Lines --- p.170 / Chapter 3.8.4 --- Effect of TNF on the Release of Calcium in Resistant Cell Lines --- p.178 / Chapter 3.8.5 --- Effect of TNF on Cell Cycle in Resistant Cell Lines --- p.185 / Chapter Chapter 4. --- General Discussion --- p.187 / Chapter 4.1 --- Tumor Necrosis Factor Induced Apoptosis in L929 Cells --- p.188 / Chapter 4.2 --- Tumor Necrosis Factor Increased the Release of Reactive Oxygen Species in L929 Cells --- p.189 / Chapter 4.3 --- Tumor Necrosis Factor Increased the Release of Calcium in L929 Cells --- p.194 / Chapter 4.4 --- Calcium Induced Reactive Oxygen Species Release in TNF- Treated L929 Cells --- p.197 / Chapter 4.5 --- Tumor Necrosis Factor Did Not Change the pH and Mitochondrial Membrane Potential in TNF-Treated L929 Cells --- p.198 / Chapter 4.6 --- Tumor Necrosis Factor Did Not Increase the Release of Reactive Oxygen Species or Calcium in Resistant Cell Lines --- p.201 / Chapter Chapter 5. --- Future Perspective --- p.204 / Chapter 5.1 --- The Relationship Between Tumor Necrosis Factor and Cytochrome c --- p.205 / Chapter 5.2 --- The Relationship Between Tumor Necrosis Factor and Mitochondrial DNA Damage --- p.206 / Chapter 5.3 --- Clinical studies with Tumor Necrosis Factor --- p.206 / References --- p.208

by Chan Hung Chuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 143-151). / Abstract also in Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / Abstract (Chinese version) --- p.v / Abbreviations --- p.viii / Content --- p.ix / Chapter CHAPTER ONE --- INTRODUCTION / Chapter 1. --- Free radical and free radical-mediated antitumor drugs --- p.1 / Chapter 2. --- Mitomycin C (MC) / Chapter 2.1 --- Drug actions of MC --- p.2 / Chapter 2.2 --- Adverse side effects of MC --- p.5 / Chapter 3. --- Doxorubicin (DOX) / Chapter 3.1 --- Drug actions of DOX --- p.7 / Chapter 3.2 --- Adverse side effects of DOX --- p.8 / Chapter 4. --- Antioxidants --- p.14 / Chapter 5. --- Effects of exogenous ATP on the antitumor activity of Doxorubicin and Mitomycin C / Chapter 5.1 --- Glutathione (GSH) and related enzymes --- p.17 / Chapter 5.2 --- Glutathione (GSH) and Anticancer Quinones --- p.19 / Chapter 5.3 --- Glutathione and the cardiac toxicity of the anticancer drugs --- p.20 / Chapter 5.4 --- Glutathione depletion in tumor cells by exogenous ATP --- p.21 / Chapter 6. --- Aim of research --- p.24 / Chapter CHAPTER TWO --- THE EFFECT OF ANTIOXIDANTS ON DOXORUBICIN- OR MITOMYCIN C-INDUCED CYTOTOXICITY ON HUMAN TUMOR AND NORMAL CELL LINES / Chapter 2.1 --- Introduction --- p.26 / Chapter 2.2 --- Materials and Methods --- p.28 / Chapter 2.3 --- Results --- p.36 / Chapter 2.4 --- Discussion --- p.60 / Chapter CHAPTER THREE --- STUDY OF CARDIOPROTECTIVE EFFECTS OF ANTIOXIDANTS AGAINST DOXORUBICIN- OR MITOMYCIN C-INDUCED TOXICITY BY LANGENDORFF PERFUEED ISOLATED RAT HEART MODEL / Chapter 3.1 --- Introduction --- p.64 / Chapter 3.2 --- Materials and Methods --- p.67 / Chapter 3.3 --- Results --- p.75 / Chapter 3.4 --- Discussion --- p.76 / Chapter CHAPTER FOUR --- THE EFFECT OF ANTIOXIDANTS DURING CHEMOTHERAPY OF DOXORUBICIN OR MITOMYCIN C IN TUMOR-BEARING MICE / Chapter 4.1 --- Introduction --- p.78 / Chapter 4.2 --- Materials and Methods --- p.80 / Chapter 4.3 --- Results --- p.83 / Chapter 4.4 --- Discussion --- p.93 / Chapter CHAPTER FIVE --- HISTOLOGICAL STUDY AND LIPID PEROXIDATION STUDY OF PROTECTIVE EFFECT OF ANTIOXIDANTS IN TUMOR-BEARING MICE TREATED WITH DOXORUBICIN OR MITOMYCIN C / Chapter 5.1 --- Introduction --- p.95 / Chapter 5.2 --- Materials and Methods --- p.98 / Chapter 5.3 --- Results --- p.103 / Chapter 5.4 --- Discussion --- p.117 / Chapter CHAPTER SIX --- EFFECT OF EXOGENOUS ATP ON THE ANTITUMOR ACTIVITY OF DOXORUBICIN AND MITOMYCIN C ON CULTURED HUMAN HEPATOMA CELLS / Chapter 6.1 --- Introduction --- p.122 / Chapter 6.2 --- Materials and Methods --- p.124 / Chapter 6.3 --- Results --- p.126 / Chapter 6.4 --- Discussion --- p.136 / Chapter CHAPTER SEVEN --- CONCLUSION / Chapter 7.1 --- Conclusion --- p.139 / Chapter 7.2 --- Future perspective --- p.141 / Bibliography --- p.142

Submitted by Geyciane Santos (geyciane_thamires@hotmail.com) on 2015-06-09T15:53:14Z No. of bitstreams: 1 Dissertação - Eleonora Dantas Dias.pdf: 1606006 bytes, checksum: 39bc4387f7de748f5319e9835177b21c (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-06-09T19:53:00Z (GMT) No. of bitstreams: 1 Dissertação - Eleonora Dantas Dias.pdf: 1606006 bytes, checksum: 39bc4387f7de748f5319e9835177b21c (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-06-09T19:55:09Z (GMT) No. of bitstreams: 1 Dissertação - Eleonora Dantas Dias.pdf: 1606006 bytes, checksum: 39bc4387f7de748f5319e9835177b21c (MD5) / Made available in DSpace on 2015-06-09T19:55:09Z (GMT). No. of bitstreams: 1 Dissertação - Eleonora Dantas Dias.pdf: 1606006 bytes, checksum: 39bc4387f7de748f5319e9835177b21c (MD5) Previous issue date: 2008-12-02 / Não Informada / The Acquired Immunodeficiency Syndrome (AIDS) constitutes a sub-epidemic in Brazil. Due to the increasing number of women infected by the virus, vertical transmission increasing substantially, and the lack of adequate prophylactic treatment, many children are infected and show manifestations of the disease in early ages. Multiples are the systems affected by the HIV virus, being frequently the skin the first organ to be affected. Skin disorders are the most common manifestation in children with HIV, being sometimes persistent and recurrent. The study objective is to analyze clinic, dermatologic and epidemiological profiles of children carriers of the virus in the City of Manaus aiming to identify the most frequent dermatosis that affect children carriers of HIV virus in the Amazon Region as to try to related these dermatosis with immunologic deterioration in this population. A study took place, where children carriers of HIV virus from the Fundação de Medicina Tropical were studied from March 2007 to July 2008. These children were submitted to dermatological examination viewing to identify dermatosis and also submitted to laboratorial exams such as viral load dosage, CD4+, and CD8+. Specific exams were also realized whenever necessary to give support to dermatosis diagnosis such as direct and culture mycological exam, bacteriologic and histopathological exams. The data collect were stored in data base elaborated using Excel and analyzed using Epi-Info, Windows versions. During the study period, 70 children HIV + were examined, all of them had AIDS and had being contaminated by vertical transmission. The media of children attended was 1.67 and the media of dermatosis by children was 1.73 dermatosis/children and among the 70 youngest, 95.5% had at least one dermatosis during the study period. The most frequent manifestation was atopic dermatitis (22.9%), prurigo estrofulo (20.0%) and wartz (18.6%). There was no statistical difference between the children in the group using HAART and the group that wasn’t using it. / A Síndrome da Imunodeficiência Adquirida (AIDS) tem se configurado como uma sub-epidemia no Brasil, devido ao crescente número de mulheres infectadas pelo vírus, a transmissão vertical aumentou significativamente, e devido a falta de tratamento profilático adequado, muitas crianças são infectadas e convivem com as manifestações da doença precocemente. Múltiplos são os sistemas acometidos pelo vírus HIV, sendo a pele muitas vezes o primeiro órgão acometido. As desordens cutâneas são as manifestações mais comuns em crianças com HIV, sendo por vezes persistentes e recorrentes. O estudo teve por objetivo analisar o perfil clínico-dermatológico e imunológico das crianças portadoras do vírus HIV na cidade de Manaus com a finalidade de identificar as dermatoses mais freqüentes que acometem as crianças portadoras do vírus HIV na população estudada, bem como tentar relacionar essas dermatoses com a deterioração do sistema imunológico dessa população. Realizou-se um estudo onde foram acompanhadas no período de março de 2007 a julho de 2008, crianças portadoras do vírus HIV atendidas na Fundação de Medicina Tropical do Amazonas. Estas crianças foram submetidas a exame dermatológico para identificação de dermatoses e a exames laboratoriais como dosagem de carga viral, CD4+, CD8+. Também foram realizados exames específicos quando se fez necessário para suporte ao diagnóstico de dermatoses como exame micológico direto e cultura, bacteriológicos e histopatológicos. Os dados coletados foram armazenados em base de dados elaborada no programa Excel e analisados no programa Epi-Info versão Windows. Durante o período estudado, foram atendidas 70 crianças HIV+, todas já apresentavam AIDS e tinham sido contaminadas por transmissão vertical. A média de consultas por criança foi de 1,67 e a média de dermatose por criança foi de 1,73 dentre os 70 menores, 95,5% apresentou pelo menos uma dermatose no período do estudo. A manifestação mais freqüente encontrada foi à dermatite atópica (22,9%), prurigo estrófulo (20%) e verruga (18,6%). Não houve diferença estatística em relação às dermatoses entre o grupo de crianças que estava em uso de Terapia antiretroviral (TARV) e o que não estava.

Introdução: A Agamaglobulinemia, a Imunodeficiência Comum Variável (IDCV) e a Síndrome Hiper IgM (SHIGM) são imunodeficiência primárias predominantemente humorais que se beneficiam da reposição de imunoglobulina humana, com redução da morbimortalidade. No entanto, apesar da reposição adequada de imunoglobulina, complicações pulmonares podem ocorrer, influenciando o prognóstico destes pacientes. Objetivo: O objetivo do estudo foi descrever as alterações morfológicas e funcionais pulmonares em pacientes com hipogamaglobulinemia primária em tratamento com reposição de imunoglobulina humana. Métodos: Foram avaliados 30 pacientes (agamaglobulinemia n=14; IDCV n=9; SHIGM n=7) que receberam imunoglobulina e antibioticoterapia profilática regularmente. A avaliação utilizou dados dos prontuários sobre o início e a evolução da doença, bem como dados de espirometria e tomografia computadorizada de tórax. O escore de Bhalla foi aplicado à tomografia mais recente de cada um dos pacientes para correlacionar as alterações tomográficas pulmonares com os dados clínicos, resultados das espirometrias e ocorrência de processos infecciosos sino-pulmonares após o início da reposição de imunoglobulina. Para as análises estatísticas utilizou-se o programa SPSS 13.0, e valores de p<0,05 foram considerados estatisticamente significantes. As variáveis nominais foram comparadas através do teste de Fisher e as contínuas, através de testes não paramétricos (Mann-Whitney, Kruskal- Wallis e Wilcoxon). Para as correlações do escore de Bhalla com as demais variáveis foi utilizado o coeficiente de Spearman. Resultados: Houve diminuição na frequência de pneumonias (p<0,001) e aumento na frequência de sinusites (p<0,001) após o início da reposição de imunoglobulina. Distúrbios ventilatórios foram evidenciados em 14 dos 23 pacientes que puderam realizaram espirometria (7 obstrutivos, 5 restritivos e 2 inconclusivos). Pacientes com bronquiectasias ao diagnóstico e aqueles à primeira avaliação tomográfica apresentaram mediana de idade mais elevada ao diagnóstico (p=0,015 e p=0,001, respectivamente) e tempo mais prolongado entre o início dos sintomas e o diagnóstico que aqueles sem bronquiectasias (p=0,010 e p=0,001, respectivamente). Sete pacientes desenvolveram bronquiectasias durante o tratamento. Pacientes com bronquiectasias à avaliação final apresentaram maior frequência de sinusites antes do início da reposição de imunoglobulina que aqueles sem bronquiectasias (p=0,010). Houve correlação estatisticamente significante do escore de Bhalla com VEF1 pré e pós-broncodilatador (r= -0,778 e r= -0,837, respectivamente), CVF (r= -0,773), FEF25-75% (r= -0,571) e com a frequência de pneumonias após o início do tratamento (r= 0,561). Conclusões: O tratamento com reposição regular de imunoglobulina e antibioticoterapia profilática reduziu a frequência e gravidade das infecções pulmonares, porém não evitou a ocorrência de sinusites, o aparecimento de bronquiectasias nem de outras alterações morfológicas e funcionais pulmonares / Introduction: Agammaglobulinemia, Common Variable Immunodeficiency (CVID) and Hyper IgM Syndrome (HIGM) are predominantly antibody deficiencies that benefit from immunoglobulin replacement therapy, with reduction of their morbidity and mortality. Despite regular immunoglobulin replacement, pulmonary complications may occur in those patients, affecting their prognosis. Objective: The aim of this study was to describe tomographic and functional pulmonary abnormalities in patients with primary hypogammaglobulinemia receiving immunoglobulin replacement therapy. Methods: Thirty patients (agammaglobulinemia n=14, CVID n=9, HIGM n=7) receiving antimicrobial prophylaxis and regular immunoglobulin infusions were evaluated. Clinical records were reviewed to obtain data concerning the onset and evolution of the disease and the results of spirometry and computed tomography of the chest. Bhalla score was applied to the most recent tomography of each patient to correlate tomographic pulmonary abnormalities with clinical data, spirometry results and the occurrence of sinusal and pulmonary infections after the onset of the immunoglobulin replacement. Statistical analysis was performed using the software SPSS 13.0 and p values < 0.05 were interpreted as statistically significant. Nominal variables were tested using Fisher´s exact test and continuous variables were tested using non-parametric tests (Mann-Whitney, Kruskal-Wallis e Wilcoxon). Spearman coefficient was used to correlate Bhalla score with the other variables. Results: The frequency of pneumonias decreased (p<0.001) and the frequency of sinusitis increased (p<0.001) after the onset of immunoglobulin replacement. Pulmonary function was abnormal in 14 of 23 patients (7 obstructive, 5 restrictive, 2 inconclusive). Patients with bronchiectasis at diagnosis and those with bronchiectasis at the first tomographic evaluation presented higher median age at diagnosis (p=0.015 and p=0.001, respectively) and longer duration between the onset of symptoms and diagnosis than those without bronchiectasis (p=0.010 e p=0.001, respectively). Seven patients developed bronchiectasis during treatment. Patients with bronchiectasis at the last tomographic evaluation presented a higher frequency of sinusitis before therapy onset than those without bronchiectasis (p=0.001). Statistically significant correlation was found between Bhalla score and pre and post bronchodilator FEV1 (r= -0.778 and r= -0.837, respectively), FVC (r= -0.773) and FEF25-75% (r= -0.571) and between Bhalla score and the frequency of pneumonias after the onset of immunoglobulin replacement therapy (r=0.561). Conclusions: Immunoglobulin replacement therapy and antimicrobial prophylaxis reduced the frequency and severity of pulmonary infections but did not prevent the occurrence of sinusitis, the development of bronchiectasis or other morphological and functional pulmonary abnormalities