Diss. (sammanfattning) Linköping : Univ. / Härtill 6 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2001. / Härtill 7 uppsatser.

Liu Kin Hang. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 88-98). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / Abstract (in Chinese) --- p.iii / Acknowledgement --- p.v / Table of Contents --- p.vi / List of Tables --- p.ix / List of Figures --- p.x / Glossary of Terms and Abbreviations --- p.xi / Chapter Section I: --- Overview --- p.1 / Chapter Chapter 1: --- Introduction --- p.2 / Chapter 1.1 --- Asthma and Assessment of A irway Inflammation --- p.2 / Chapter 1.1.1 --- Assessment of A irway Inflammation --- p.4 / Chapter 1.1.2 --- Invasive and Noninvasive Methods --- p.4 / Chapter 1.1.3 --- Exhaled Nitric Oxide as a Diagnostic Marker and Its Correlation with Other Markers of Inflammation --- p.6 / Chapter 1.1.4 --- Normal Reference Studies of Exhaled Nitric Oxide --- p.8 / Chapter 1.2 --- Aim of Study --- p.10 / Chapter Chapter 2: --- Plan of Study --- p.11 / Chapter Section II: --- Literature Review --- p.13 / Chapter Chapter 3: --- Nitric Oxide Biology --- p.15 / Chapter 3.1 --- Exhaled Nitric Oxide Production in Airway --- p.15 / Chapter 3.2 --- Nitric Oxide Production and Function --- p.16 / Chapter 3.3 --- Nitric Oxide Synthase Pathway --- p.18 / Chapter 3.4 --- Factors Affecting Exhaled Nitric Oxide Level --- p.21 / Chapter 3.4.1 --- Procedure-related Factors --- p.22 / Chapter 3.4.1.1 --- Nasal Nitric Oxide Contamination --- p.22 / Chapter 3.4.1.2 --- Exhalation Procedure 226}0´ؤؤStarting Lung Volumes --- p.23 / Chapter 3.4.1.3 --- Exhalation Procedure 226}0ؤ Flow --- p.23 / Chapter 3.4.1.4 --- Circadian Rhythm --- p.25 / Chapter 3.4.2 --- Patient Factors --- p.26 / Chapter 3.4.2.1 --- Sex --- p.26 / Chapter 3.4.2.2 --- Upper Respiratory Tract Infection --- p.26 / Chapter 3.4.2.3 --- Diet and Exhaled Nitric Oxide --- p.27 / Chapter 3.4.2.4 --- Effect of Spirometry and Exercise --- p.28 / Chapter 3.4.3 --- Environmental Factors --- p.28 / Chapter Chapter 4: --- Exhaled Nitric Oxide in Asthmatics and Its Relationship to Anti-inflammatory Treatment --- p.31 / Chapter Chapter 5: --- Relationship of Exhaled Nitric Oxide with Other Inflammatory Markers --- p.33 / Chapter 5.1 --- Correlation of Findings from Biopsy and Bronchoalveolar Lavage with Exhaled Nitric Oxide --- p.33 / Chapter 5.2 --- "Exhaled Nitric Oxide, Induced Sputum Analysis and Sputum Eosinophil Cationic Protein" --- p.35 / Chapter Section III: --- Original Study --- p.37 / Chapter Chapter 6: --- Methodology --- p.38 / Chapter 6.1 --- Study Population --- p.38 / Chapter 6.2 --- The International Study of Asthma and Allergies in Childhood --- p.40 / Chapter 6.3 --- ISAAC Questionnaires --- p.42 / Chapter 6.4 --- Standardized Approach for Answering Questions in the Field --- p.44 / Chapter 6.5 --- Anthropometric Measurements --- p.45 / Chapter 6.6 --- Exhaled Nitric Oxide Measurement --- p.46 / Chapter 6.6.1 --- "NIOY® (Aerocrine AB, Stockholm, Sweden)" --- p.46 / Chapter 6.6.2 --- Calibration Procedures --- p.47 / Chapter 6.6.3 --- Exhaled Nitric Oxide Measurement --- p.48 / Chapter 6.7 --- Classification of Subjects --- p.51 / Chapter 6.8 --- Statistical Analysis --- p.53 / Chapter Chapter 7: --- Results --- p.54 / Chapter 7.1 --- Subjects and Demography --- p.54 / Chapter 7.2 --- Exhaled Nitric Oxide in Chinese Children --- p.58 / Chapter 7.3 --- Exhaled Nitric Oxide in Caucasians and Other Ethnic Groups --- p.66 / Chapter Chapter 8: --- Discussion --- p.69 / Chapter Chapter 9: --- Conclusion and Further Studies --- p.76 / Appendix 1 Questionnaires (Chinese Version) --- p.80 / Appendix 2 Questionnaires (English Version) --- p.84 / References --- p.88

Lau Yin Kei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 58-79). / Abstracts in English and Chinese. / Acknowledgement --- p.I / Abstract --- p.IV / Abstract in Chinese --- p.VI / Abbreviations --- p.VIII / Introduction --- p.1 / Chapter 1.1 --- Prevalence of COPD and asthma in Hong Kong --- p.1 / Chapter 1.2 --- Players in pathogenesis of COPD --- p.2 / Chapter 1.3 --- Players in pathogenesis of asthma --- p.4 / Chapter 1.4 --- The use of exhaled breath condensate in previous studies --- p.6 / Chapter 1. 5 --- Brief overview of chemokines --- p.8 / Chapter 1.6 --- Objective of this study --- p.12 / Materials and methods --- p.14 / Chapter 2.1 --- Study population --- p.14 / Chapter 2.1.1 --- Patients with COPD and control subjects --- p.14 / Chapter 2.1.2 --- Patients with asthma and control subjects --- p.15 / Chapter 2.2 --- Lung function --- p.15 / Chapter 2.3 --- Dyspnoea score measurement of patients with COPD --- p.16 / Chapter 2.4 --- Classification of patients and asthma severity --- p.16 / Chapter 2.5 --- Skin prick test and blood tests --- p.16 / Chapter 2.6 --- Collection of exhaled breath condensate --- p.17 / Chapter 2.7 --- Measurement of constituent in EBC --- p.17 / Chapter 2.7.1 --- "Measurement of 8-isoprostane, MCP-1 and GROα in patients with COPD and the corresponding control subjects" --- p.17 / Chapter 2.7.2 --- Measurement of eotaxin and MDC of patients with asthma and the corresponding control subjects --- p.18 / Chapter 2.8 --- Reproducibility of exhaled breath constituent --- p.18 / Chapter 2.8.1 --- "Assessment of reproducibility of the exhaled MCP-1, GROα and8- isoprostane measurements" --- p.19 / Chapter 2.8.2 --- Assessment of reproducibility of the exhaled eotaxin and MDC measurement --- p.19 / Chapter 2.9 --- Statistical analysis --- p.19 / Results --- p.21 / Chapter 3.1 --- Patients with COPD and corresponding control subjects --- p.21 / Chapter 3.2 --- Patients with asthma and corresponding control subjects --- p.28 / Discussion --- p.36 / Chapter 4.1 --- "Exhaled 8-isoprostane, GRO-α and MCP-1 of patients with COPD and corresponding control subjects" --- p.36 / Chapter 4.2 --- Exhaled eotaxin and MDC from patients with asthma and corresponding control subjects --- p.43 / Chapter 4.3 --- Technical aspects of EBC assessment --- p.49 / Future prospect --- p.54 / Conclusion --- p.56 / References --- p.58 / Tables and Figures / Table 1. Demographics of the COPD and control subjects --- p.22 / Figure 1. The level of 8-isoprostane in the exhaled breath condensate of COPD and control subjects --- p.23 / Figure 2. The level of GROa in the exhaled breath condensate of COPD and control subjects --- p.25 / "Figure 3 Bland and Altman's Plot of the repeatability of 8-isoprostane, GROa and MCP-1 in the exhaled breath condensate of normal controls" --- p.27 / Table2. Clinical and physiological details of the subjects --- p.29 / Figure 4. Level of eotaxin in exhaled breath condensate of asthma and control subjects --- p.30 / Figure 5 Level of MDC in exhaled breath condensate of asthma and control subjects --- p.31 / Table 3. Levels of eotaxin and MDC in exhaled breath condensate of asthma subjects on different dose of inhaled corticosteroids --- p.33 / Figure 6. Relationship between exhaled breath condensate level of MDC and total serum IgE level --- p.35

O infarto agudo do miocárdio (IAM) é definido como uma necrose do miocárdio resultante de um comprometimento agudo de sua irrigação sang?ínea. As manifestações de insuficiência cardíaca (ICC) são comuns em pacientes com doença arterial coronariana (DAC) aguda ou crônica, acarretando significativa morbidade e mortalidade. O objetivo foi avaliar a função global e regional do ventrículo esquerdo (VE), através da Ressonância Magnética (IRM) com a técnica dos marcadores miocárdicos na evolução clínica dos pacientes na fase tardia do infarto da parede anterior do VE. Foi realizado seguimento longitudinal prospectivo da evolução da função da parede do ventrículo esquerdo. Foram avaliadas e comparadas entre si as várias etapas evolutivas de pacientes encaminhados pela Unidade Clínica de Coronariopatia Crônica e Unidade de Cirurgia Torácica e Cardiovascular do Incor-FMUSP. Entre dezembro de 2000 e fevereiro de 2003, estudamos 24 pacientes (19 homens e cinco mulheres), idade média de 54,33 ± 10,11 anos. Os estudos foram realizados na inclusão do paciente no protocolo, após 4 meses e 10 meses de seguimento. Os estudos foram realizados em repouso (rep) e durante o estímulo inotrópico com baixa dose de dobutamina (dob) (10 mcg/kg/ml). Nenhum paciente desenvolveu sintomas durante a infusão de dobutamina. Foram estudados os volumes diastólico (VDF) e sistólico (VSF) finais e a fração de ejeção (FE) com a técnica de cine ressonância, utilizando-se o método de Simpson para a análise. A função global e regional foi analisada com a técnica dos marcadores miocárdicos através da análise do encurtamento circunferencial (EC) global e regional nas áreas remotas, adjacentes e com infarto. Os resultados mostraram estabilidade nos valores encontrados (VDF,VSF e FE), tanto em repouso como durante a infusão de dobutamina (p=NS). A análise da função do VE com a técnica dos marcadores miocárdicos no grupo clínico mostrou melhora significante nos exames de controle, após 4 meses e 10 meses de seguimento na comparação das médias globais (p<0,001). Na comparação entre o repouso e a infusão de dobutamina do exame 1 (p=0,01), no exame 2 (p<0,001), no exame 3 (p<0,001). Nas regiões com infarto não houve diferença significante entre o grupo 1, 2 e 3 (comprometimento mural <=75%), o que pode ser evidenciado entre os grupos 1, 2 e 3 vs. grupo 4 ( comprometimento mural >75%)(p<0,001). Em conclusão, este estudo mostra a manutenção das variáveis da função ventricular esquerda (volumes e fração de ejeção) durante a evolução clínica. Os pacientes em acompanhamento clínico avaliados com a técnica dos marcadores miocárdicos mostraram melhora quantitativa da função global e regional na área com infarto, o que indica que este índice pode ser mais sensível na avaliação evolutiva da função ventricular esquerda. / Acute myocardial infarction is defined as a myocardial necrosis resultant from an acute comitment of blood irrigation. Manifestations of cardiac failure are common in patients with acute or cronic coronary artery disease, with significant morbidity and mortality. Our goal was to valuate clinic evolution in patients with chronic myocardial infarction on the global and regional left ventricle (LV) systolic function by magnetic resonance imaging (MRI) with myocardial tagging technique. A longitudinal follow-up of LV function was done in patients on clinical treatment. Patients were referred from the Coronary Unit or Thoracic and Cardiovascular Surgery of the Heart Institute (InCor) - University of São Paulo Medical School. From December 2000 to February 2003, we studied 24 patients (19 men), mean age 54.33 + 10.11 years. Medical group was evaluated and compared among each follow-up steps (at protocol inclusion (E1), after 4 months (E2) and 10 months (E3) follow-up. The studies were performed at rest (R) and during inotropical stimulus with low dose of dobutamine (D) (10 mcg/kg/ml). The end diastolic (EDV) and systolic (ESV) volumes were studied and ejection fraction (EF) with the cine resonance technique using Simpson method. Global and regional function was also analized with the myocardial tagging technique. The variables studied were the global and segmental circumferencial shortening (CS) in remote, adjacent and infarcted areas. Medical group showed no differences in EDV, ESV and EF (p=NS). LV function analysis with the myocardial tagging techniques showed an significant improvement in the exams performed at E1, E2 and E3 follow-up on global average (p<0,001), in E1 at R vs. D (p=0,023), E2 R vs. D (p=0,001), E3 R vs. D (p=0,008). CS in infarcted areas showed no significant differences between group 1 , 2 and 3 (infarcted area <=75%) but differences were seen between group 1 , 2 and 3 vs. group 4 (infarcted area >75%)(p=0,006). In conclusion, this study showed no differences in LV volumes and ejection fraction. The patients in clinical follow up showed quantitative improvement at global and regional function at infarcted areas with the myocardial tagging techniques, which seen a better index for LV function follow-up

As miocardiopatias dilatadas representam 87% das miocardiopatias e apresentam evolução adversa com grande morbi-mortalidade. Vários marcadores de prognóstico são bem definidos, entretanto, um marcador estrutural se faz necessário. Estudamos através da realização da biópsia endomiocárdica e exame ecocardiográfico, 9 indivíduos sem doença estrutural miocárdica (controle) e 45 pacientes com miocardiopatia dilatada grave de etiologia idiopática (MCDI) e chagásica (MCDC). Observamos se havia relação entre a quantidade de colágeno miocárdico intersticial (FVCI) e a sobrevida destes pacientes, se a FVCI diferia entre as etiologias, e se a fibrose interferia na função e geometria do miocárdio. Observamos que a FVCI foi 15x maior nos miocardiopatas em relação ao grupo controle, mas não diferiu em relação às MCDI e MCDC (FVCI % MCDC = 6,83 ± 5,47; MCDI = 5,75 ± 4,45; controle = 0,42 ± 0,14*; p<0,001). Não houve relação da FCVI com a sobrevida dos pacientes com miocardiopatias (MCDI-FVCI £5,53 (20,0%) ou >5,53 (0,0%) (p=0,249), e na MCDC-FVCI £5,53 (0,0%) ou >5,53 (7,7%) (p=0,587) e apenas na MCDI a fração de ejeção do ventrículo esquerdo (FEVE) teve relação com a FVCI. O diâmetro diastólico final do ventrículo esquerdo não se correlacionou com a FCVI nas duas etiologias. Conclusão: a fibrose miocárdica não diferiu entre as duas etiologias, não se correlacionou com o prognóstico das MCDC e MCDI e apenas na MCDI ela se correlacionou com a FEVE. / Dilated cardiomyopathies represent 87% of all cardiomyopathies and they have adverse prognosis with high morbidity and mortality. There are several prognostic markers, however, a structural one has not been described yet. Seems to be very important to find out whether morphological changes upon myocardial structure would affect the prognosis. We studied, using endomyocardial biopsy and 2D-echocardiogram, 9 patients with no structural myocardial changes (control) and 45 patients with severe dilated cardiomyopathies. They were divided according the etiology of cardiomyopathy into idiopathic group (IDCM) or Chagas group (CDCM). We analyzed the correlation between interstitial myocardial collagen (ICVF) and survival rates. We also evaluated the difference of ICVF between these groups and whether it correlates with geometric and functional changes of the heart. We observed that ICVF was 15 times higher in cardiomyopathies patients than in control group, but it did not differ between CDCM and IDCM (ICVF% CDCM = 6.83 ± 5.47; IDCM = 5.75 ± 4.45; control = 0.42 ± 0.14*; p<0.001). The ICVF did not correlate to survival rate in cardiomyopathies patients (IDCM-ICVF £5.53 (20.0%) or >5.53 (0.0%) (p=0.249), and CDCM-ICVF £5.53 (0.0%) or >5.53 (7.7%) (p=0.587). We observed a significant correlation between ICVF and left ventricular ejection fraction (LVEF) only on DMC, the ICVF did not correlate to left ventricular diastolic diameter in either etiology. Conclusion: the myocardial fibrosis did not differ between these two etiologies, it did not correlate to prognosis either in the IDCM or CDCM and only in the IDCM the ICVF correlated to the LVEF.

O infarto agudo do miocárdio (IAM) é definido como uma necrose do miocárdio resultante de um comprometimento agudo de sua irrigação sang?ínea. As manifestações de insuficiência cardíaca (ICC) são comuns em pacientes com doença arterial coronariana (DAC) aguda ou crônica, acarretando significativa morbidade e mortalidade. O objetivo foi avaliar a função global e regional do ventrículo esquerdo (VE), através da Ressonância Magnética (IRM) com a técnica dos marcadores miocárdicos na evolução clínica dos pacientes na fase tardia do infarto da parede anterior do VE. Foi realizado seguimento longitudinal prospectivo da evolução da função da parede do ventrículo esquerdo. Foram avaliadas e comparadas entre si as várias etapas evolutivas de pacientes encaminhados pela Unidade Clínica de Coronariopatia Crônica e Unidade de Cirurgia Torácica e Cardiovascular do Incor-FMUSP. Entre dezembro de 2000 e fevereiro de 2003, estudamos 24 pacientes (19 homens e cinco mulheres), idade média de 54,33 ± 10,11 anos. Os estudos foram realizados na inclusão do paciente no protocolo, após 4 meses e 10 meses de seguimento. Os estudos foram realizados em repouso (rep) e durante o estímulo inotrópico com baixa dose de dobutamina (dob) (10 mcg/kg/ml). Nenhum paciente desenvolveu sintomas durante a infusão de dobutamina. Foram estudados os volumes diastólico (VDF) e sistólico (VSF) finais e a fração de ejeção (FE) com a técnica de cine ressonância, utilizando-se o método de Simpson para a análise. A função global e regional foi analisada com a técnica dos marcadores miocárdicos através da análise do encurtamento circunferencial (EC) global e regional nas áreas remotas, adjacentes e com infarto. Os resultados mostraram estabilidade nos valores encontrados (VDF,VSF e FE), tanto em repouso como durante a infusão de dobutamina (p=NS). A análise da função do VE com a técnica dos marcadores miocárdicos no grupo clínico mostrou melhora significante nos exames de controle, após 4 meses e 10 meses de seguimento na comparação das médias globais (p<0,001). Na comparação entre o repouso e a infusão de dobutamina do exame 1 (p=0,01), no exame 2 (p<0,001), no exame 3 (p<0,001). Nas regiões com infarto não houve diferença significante entre o grupo 1, 2 e 3 (comprometimento mural <=75%), o que pode ser evidenciado entre os grupos 1, 2 e 3 vs. grupo 4 ( comprometimento mural >75%)(p<0,001). Em conclusão, este estudo mostra a manutenção das variáveis da função ventricular esquerda (volumes e fração de ejeção) durante a evolução clínica. Os pacientes em acompanhamento clínico avaliados com a técnica dos marcadores miocárdicos mostraram melhora quantitativa da função global e regional na área com infarto, o que indica que este índice pode ser mais sensível na avaliação evolutiva da função ventricular esquerda. / Acute myocardial infarction is defined as a myocardial necrosis resultant from an acute comitment of blood irrigation. Manifestations of cardiac failure are common in patients with acute or cronic coronary artery disease, with significant morbidity and mortality. Our goal was to valuate clinic evolution in patients with chronic myocardial infarction on the global and regional left ventricle (LV) systolic function by magnetic resonance imaging (MRI) with myocardial tagging technique. A longitudinal follow-up of LV function was done in patients on clinical treatment. Patients were referred from the Coronary Unit or Thoracic and Cardiovascular Surgery of the Heart Institute (InCor) - University of São Paulo Medical School. From December 2000 to February 2003, we studied 24 patients (19 men), mean age 54.33 + 10.11 years. Medical group was evaluated and compared among each follow-up steps (at protocol inclusion (E1), after 4 months (E2) and 10 months (E3) follow-up. The studies were performed at rest (R) and during inotropical stimulus with low dose of dobutamine (D) (10 mcg/kg/ml). The end diastolic (EDV) and systolic (ESV) volumes were studied and ejection fraction (EF) with the cine resonance technique using Simpson method. Global and regional function was also analized with the myocardial tagging technique. The variables studied were the global and segmental circumferencial shortening (CS) in remote, adjacent and infarcted areas. Medical group showed no differences in EDV, ESV and EF (p=NS). LV function analysis with the myocardial tagging techniques showed an significant improvement in the exams performed at E1, E2 and E3 follow-up on global average (p<0,001), in E1 at R vs. D (p=0,023), E2 R vs. D (p=0,001), E3 R vs. D (p=0,008). CS in infarcted areas showed no significant differences between group 1 , 2 and 3 (infarcted area <=75%) but differences were seen between group 1 , 2 and 3 vs. group 4 (infarcted area >75%)(p=0,006). In conclusion, this study showed no differences in LV volumes and ejection fraction. The patients in clinical follow up showed quantitative improvement at global and regional function at infarcted areas with the myocardial tagging techniques, which seen a better index for LV function follow-up

As miocardiopatias dilatadas representam 87% das miocardiopatias e apresentam evolução adversa com grande morbi-mortalidade. Vários marcadores de prognóstico são bem definidos, entretanto, um marcador estrutural se faz necessário. Estudamos através da realização da biópsia endomiocárdica e exame ecocardiográfico, 9 indivíduos sem doença estrutural miocárdica (controle) e 45 pacientes com miocardiopatia dilatada grave de etiologia idiopática (MCDI) e chagásica (MCDC). Observamos se havia relação entre a quantidade de colágeno miocárdico intersticial (FVCI) e a sobrevida destes pacientes, se a FVCI diferia entre as etiologias, e se a fibrose interferia na função e geometria do miocárdio. Observamos que a FVCI foi 15x maior nos miocardiopatas em relação ao grupo controle, mas não diferiu em relação às MCDI e MCDC (FVCI % MCDC = 6,83 ± 5,47; MCDI = 5,75 ± 4,45; controle = 0,42 ± 0,14*; p<0,001). Não houve relação da FCVI com a sobrevida dos pacientes com miocardiopatias (MCDI-FVCI £5,53 (20,0%) ou >5,53 (0,0%) (p=0,249), e na MCDC-FVCI £5,53 (0,0%) ou >5,53 (7,7%) (p=0,587) e apenas na MCDI a fração de ejeção do ventrículo esquerdo (FEVE) teve relação com a FVCI. O diâmetro diastólico final do ventrículo esquerdo não se correlacionou com a FCVI nas duas etiologias. Conclusão: a fibrose miocárdica não diferiu entre as duas etiologias, não se correlacionou com o prognóstico das MCDC e MCDI e apenas na MCDI ela se correlacionou com a FEVE. / Dilated cardiomyopathies represent 87% of all cardiomyopathies and they have adverse prognosis with high morbidity and mortality. There are several prognostic markers, however, a structural one has not been described yet. Seems to be very important to find out whether morphological changes upon myocardial structure would affect the prognosis. We studied, using endomyocardial biopsy and 2D-echocardiogram, 9 patients with no structural myocardial changes (control) and 45 patients with severe dilated cardiomyopathies. They were divided according the etiology of cardiomyopathy into idiopathic group (IDCM) or Chagas group (CDCM). We analyzed the correlation between interstitial myocardial collagen (ICVF) and survival rates. We also evaluated the difference of ICVF between these groups and whether it correlates with geometric and functional changes of the heart. We observed that ICVF was 15 times higher in cardiomyopathies patients than in control group, but it did not differ between CDCM and IDCM (ICVF% CDCM = 6.83 ± 5.47; IDCM = 5.75 ± 4.45; control = 0.42 ± 0.14*; p<0.001). The ICVF did not correlate to survival rate in cardiomyopathies patients (IDCM-ICVF £5.53 (20.0%) or >5.53 (0.0%) (p=0.249), and CDCM-ICVF £5.53 (0.0%) or >5.53 (7.7%) (p=0.587). We observed a significant correlation between ICVF and left ventricular ejection fraction (LVEF) only on DMC, the ICVF did not correlate to left ventricular diastolic diameter in either etiology. Conclusion: the myocardial fibrosis did not differ between these two etiologies, it did not correlate to prognosis either in the IDCM or CDCM and only in the IDCM the ICVF correlated to the LVEF.

Recent studies have revealed extensive cortical demyelination in patients with progressive multiple sclerosis (MS). Demyelination in gray matter lesions is associated with activation of microglia. Macrophages and microglia are known to express myeloperoxidase (MPO) and generate reactive oxygen species during myelin phagocytosis in the white matter. In the present study we examined the extent of microglial activation in the cerebral cortex and the relationship of microglial activation and MPO activity to cortical demyelination. Twenty-one cases of neuropathologically confirmed multiple sclerosis, with 34 cortical lesions, were used to assess microglial activation. HLA-DR immunolabeling of activated microglia was significantly higher in demyelinated MS cortex than control cortex and, within the MS cohort, was significantly greater within cortical lesions than in matched non-demyelinated areas of cortex. In homogenates of MS cortex, cortical demyelination was associated with significantly elevated MPO activity. Immunohistochemistry revealed MPO in CD68-positive microglia within cortical plaques, particularly toward the edge of the plaques, but not in microglia in adjacent non-demyelinated cortex. Cortical demyelination in MS is associated with increased activity of MPO, which is expressed by a CD68-positive subset of activated microglia, suggesting that microglial production of reactive oxygen species is likely to be involved in cortical demyelination.

Introdução: Apesar dos avanços nas técnicas de diagnóstico e tratamento, o câncer de próstata avançado ainda é uma condição letal. Terapêuticas mais eficazes são necessárias para reduzir as taxas de morbi-mortalidade associadas à doença. A Proteína-78 regulada pela glicose (GRP78), uma proteína de choque térmico envolvida na apresentação de antígenos, foi recentemente descrita como sendo um possível marcador molecular para o câncer de próstata. Ainda mais, a resposta imune a essa proteína mostrou correlação com o desenvolvimento de doença hormônio-independente e com pior sobrevida para a doença. Objetivos: Neste estudo, avaliou-se a hipótese de que a GRP78 poderia ser usada como marcador molecular em câncer de próstata no desenvolvimento de um sistema de receptor-ligante, através do uso da tecnologia de apresentação de fagos. Casuística e métodos: Inicialmente, foram clonados dois peptídeos que apresentam afinidade à proteína regulada pela GRP78 (os peptídeos WIFPWIQL e WDLAWMFRLPVG) no vetor fUSE5, criando-se fagos com capacidade teórica de ligação à mesma proteína. Posteriormente foi testada a capacidade de ligação desses fagos à GRP78 na membrana de células prostáticas malignas em solução, em xeno-tumores in vivo e em metástases ósseas de câncer de próstata humano. Resultados: Demonstrou-se que ambos os fagos se ligam especificamente à GRP78 in vitro, em comparação à proteínas com seqüência semelhante (proteínas de choque térmico 70 e 90) e não semelhante (albumina sérica bovina). Em seguida, mostrou-se que esses fagos se ligam com afinidade pelo menos 30 vezes maior à células de câncer de próstata que o fago controle, e que os fagos são internalizados por essas células. Posteriormente, mostrou-se que os fagos rastrearam xeno-tumores prostáticos quando injetados in vivo num modelo animal de câncer de próstata. Finalmente, mostrou-se que os fagos ligam-se especificamente à GRP78 expressa em metástases ósseas de adenocarcinoma prostático humano. Conclusões: Os fagos criados apresentam capacidade de ligação específica à GRP78 in vitro, em células em suspensão e in vivo. A estratégia e o sistema de receptor-ligante definidos no presente estudo podem ter implicacões relevantes no desenvolvimento de terapias dirigidas para o tratamento do câncer de próstata. / Introduction: Despite the advances in diagnosis and treatment, advanced prostate cancer remains a lethal condition. Improved methods of therapy are needed to reduce the morbidity and mortality rates associated with this disease. The Glucose-regulated protein-78 (GRP78), a stress-responsive heat-shock protein involved in antigen presentation, was recently described as a possible molecular marker for prostate cancer. Moreover, immune response against this protein was shown to have correlation with the development of androgen-independent prostate cancer and shorter overall survival. Objectives: We hipothesized that GRP78 could be used as a molecular marker for prostate cancer in the development of a receptor-ligand system, by using phage display technology. Patients and methods: We initially cloned two GRP78-targeting peptides (WIFPWIQL and WDLAWMFRLPVG) into a fUSE5-based phage. We then tested binding capacity of the phage to GRP78 in vitro, to GRP78 expressed in intact prostate cancer cell membranes, to a prostate cancer xenograft and to human bone metastases. Results: We showed that both phage created bound specifically to GRP78 in vitro, in comparison to related (Heat-shock proteins 70 and 90) and unrelated control proteins (bovine serum albumin). Next, we showed that these phage bound at least 30 times more to prostate cancer cells than the control phage, and were also internalized into these cells. Both GRP78-binding phage showed a strong homing in vivo to a human prostate cancer xenograft in a mouse model. Finally, we showed that both phage bound specifically to GRP78 expressed in human prostate cancer bone metastases. Conclusions: Both phage are capable of binding specifically to GRP78 in vitro, in the context of intact prostate cancer cells and in vivo. The strategy and the ligand-receptor system we have defined in this study may have relevant implications in the development of targeted therapies for the treatment of prostate cancer.