The paradigm shift in cardiovascular biology has been the understanding that atherosclerosis involves not just a mechanical deposition of lipids in the vessel wall, but a dynamic process involving the inflammatory response with cellular infiltration and inflammatory mediator expression. Typical cellular elements that have been studied include endothelial cells, vascular smooth muscle, T lymphocyte and the macrophage. Recent data suggests a role for the human mast cell. The human mast cell is a tissuedwelling cell, typically perivascular in distribution. This multifunctional cell responds rapidly to challenge with the release of inflammatory mediators that can orchestrate an immune response and may have relevance to atherogenesis. Mast cells have been shown to modulate various aspects of cardiovascular disease such as atherogenesis (endothelial activation, cytokine generation and foam cell formation) as well as rupture of an unstable atheromatous plaque. Mast cell activation in the context of cardiovascular disease may occurby cognate cell-cell interactions (interactions with macrophages, T cells, endothelial cells or smooth muscle) or by non-cognate means (such as lipoproteins and other proatherogenic components). More studies are required in order to better understand the molecular role of mast cells in vascular inflammatory disease.

Objectives: To date little is known of the interaction between diabetes and sex hormones in the vasculature. A number of studies suggest that premenopausal diabetic women loose their gender based cardiovascular protection. However, there is insufficient evidence to explain the mechanism underlying the loss of this gender based cardioprotection in premenopausal diabetic women. The objectives of this study were to investigate whether there is a gender difference in the aortic endothelial function in · streptozotocin (STZ, 58 mg/kg, iv)-induced diabetic rats, and the potential role of superoxide and cyclooxygenase (COX) metabolites in diabetes-induced vascular dysfunction.

Dislipidemia é implicada no processo aterosclerótico do LES. A descrição de aLPL no LES associado a hipertrigliceridemia levou-nos a analisar esse anticorpo no contexto da inflamação envolvida na aterogênese. aLPL foi encontrado em 38% dos pacientes com LES com altos níveis de triglicérides. Correlação positiva significante foi observada entre aLPL e PCR, VHS, SLEDAI, anti-DNA, anti-cardiolipina e CH100 baixo. Análise de regressão múltipla confirmou a forte associação entre aLPL e PCR. Esses dados dão suporte à associação entre inflamação, resposta imune e dislipidemia, introduzindo o aLPL como um novo componente nos complexos eventos da aterogênese do LES / Dyslipidemia is implicated in the atherosclerosis process of SLE. The description of aLPL in SLE associated with hypertrigliceridemia prompted us to analyze this antibody in the context of the inflammation involved in the atherogenesis. aLPL was found in 38 por cento of SLE patients with high levels of triglycerides. Significant positive correlation was observed between aLPL and CRP, ESR, SLEDAI, anti-DNA, anti-cardiolipin and low CH100. Multiple regression analysis confirmed the strong association between aLPL and CRP. These data support the link between inflammation, immune response and dyslipidemia, introducing anti-LPL as new player in the complex events of atherogenesis in SLE

A hipercolesterolemia familiar (HF) é caracterizada por concentrações elevadas de LDL-c e alta prevalência de doença arterial coronária (DAC) precoce. Entretanto, o curso da DAC nos portadores de HF é variável e pode ser influenciado por outros fatores de risco. O objetivo foi avaliar parâmetros clínicos e nutricionais de adultos portadores de HF heterozigótica por estudo do tipo transversal. Coletou-se do prontuário dos pacientes resultados de exames laboratoriais, medidas de pressão arterial e diagnósticos clínicos. Verificou-se a concordância ou não entre as categorias de risco pelos escores de Framingham (ERF) e pelos critérios estabelecidos para os portadores de HF. Antecedentes pessoais e familiares para DAC, tabagismo, atividade física, consumo alimentar de gorduras, fibras e bebidas alcoólicas foram obtidos por questionário e medidas antropométricas foram aferidas. Foram comparados os grupos com e sem Síndrome Metabólica (SM) e os grupos com e sem DAC por análise univariada. Após, foram verificados os fatores determinantes para o desenvolvimento da DAC mediante modelo de regressão multivariada. Foram entrevistados 110 pacientes (68 mulheres) com média de idade de 48,9 ± 16,2 anos. A presença de história familiar de DAC precoce foi relatada por 67 (61,5%) pacientes. A hipertensão foi encontrada em 59 (53,6%), SM em 38 (34,9%), DAC em 30 (27,3%), HDL-c baixo em 28 (25,5%), diabete melito em 17 (15,5%), 25 (22,7%) eram ex-fumantes e 12 (10,9%) tabagistas. Com a comparação das categorias de risco observou-se discrepância em 77,5% dos casos entre os ERF e os critérios estabelecidos para a população de HF. Quanto ao estado nutricional, 47 (42,7%) eram pré-obesos e 61 (55,4%) com circunferência da cintura alterada. O consumo de gorduras, fibras e bebidas alcoólicas foi considerado adequado. Encontrou-se grande número de sedentários (77%). O grupo dos pacientes com SM tinha idade mais avançada (55 vs 46 anos; p = 0,002), maior número de mulheres (76,3%; p = 0,02) e portadores de DAC (42,1%; p = 0,013). O grupo dos coronarianos tinha idade mais avançada (55 vs 47 anos; p = 0,004), mais pacientes do sexo masculino (60%; p = 0,004), maior presença de hipertensos (90%; p = 0,001), exfumantes (40%; p = 0,008), com SM (53,3%; p = 0,013), HDL-c baixo (53,3%; p = 0,001) e antecedente de infarto agudo do miocárdio (IAM) em irmãos (50%; p = 0,012). As medidas antropométricas, o consumo alimentar e a atividade física não foram diferentes entre os grupos. Após análise de regressão multivariada os fatores de risco determinantes para o desenvolvimento da DAC foram HDL-c baixo (OR 8,4; IC 95% 2,7-27,6), sexo masculino (OR 7,3; IC 95% 2,1-24,7), história de IAM em irmãos (OR 3,4; IC 95% 1,1-10,5) e idade avançada (OR 1,06; IC 95% 1,02-1,1). Em nossa população, HDL-c baixo, sexo masculino, história de IAM em irmãos e idade foram fatores independentes para o desenvolvimento da DAC. / Familial hypercholesterolaemia (FH) is characterized by raised concentrations of LDL-c and high prevalence of premature coronary artery disease (CAD). However, the course of the CAD in the FH is variable and can be influenced by other risk factors. The aim of the study was to assess clinical and nutritional parameters in adults with heterozigous FH by a cross sectional study. Laboratory exams, blood pressure measurement and clinical diagnosis were collected. Agreement or not between the categories of risk by Framingham scores and for established criteria for the FH subjects was verified. Personal and familial history for CAD, smoken habit, physical activity, fats, fibers and alcohol consumption were assessed by questionnaire and anthropometric measurement were verified. The groups with and without Metabolic Syndrome (MS) and groups with and without CAD were compared by univariated analysis. After, multivaried analysis (MVA) was used to assess the significance of differences in risk factors. The sample was composed by 110 patients (68 women) with average of age of 48.9 ± 16.2 years. The presence of familial history of premature CAD was detected in 67 (61.5%)subjects. Hypertension was found in 59 (53.6%), MS in 38 (34.9%), CAD in 30 (27.3%), low HDL-c in 28 (25.5%), diabetes in 17 (15.5%), 25 (22,7%) and 12 (10,9%) were respectively former and current smokers. In the comparison of the risk categories discrepancy was observed in 77.5% of the cases between the Framingham scores and the established criteria for the FH population. Analyzing the nutritional profile, 47 (42.7%) were overweight and 61 (55.4%) had increased waist circumference. The consumption of fats, fibers and alcohol were considered satisfactory. A great number of sedentary subjects was found (77%). The patients with MS were older (55 vs. 46 years; p = 0.002), had a greater number of women (76.3%; p = 0.02) and CAD (42.1%; p = 0.013). CAD subjects were older (55 vs. 47 years; p = 0.004), had a higher prevalence of males (60%; p = 0.004), hypertension (90%; p = 0.001), former smokers(40%; p = 0.008), MS (53.3%; p = 0.013), low HDL-c (53.3%; p = 0.001) and history of myocardial infarction in brothers (50%; p = 0.012). There were no differences between the groups regarding anthropometric measurements, consumption of fats, fiber and alcohol and physical activity. After MVA, independent risk factors for CAD were low HDL-c (OR 8.4; CI 95% 2.7-27.6), male gender (OR 7.3; CI 95% 2.1-24.7), history of myocardial infarction in brothers (OR 3.4; CI 95% 1.1-10.5) and advanced age (OR 1.06; CI 95% 1.02-1.1). In our population, low HDL-c, male gender, history of myocardial infarction in brothers and age were independently associated with the risk of CAD.

Objetivos: Muitas questões permanecem sobre as causas da aterosclerose acelerada nos pacientes com doenças inflamatórias sistêmicas como a artrite reumatóide (AR). Estudos na população geral sugeriram que além da inflamação existe uma participação patogênica da auto-imunidade na aterosclerose e discutem a possível associação dos anticorpos contra fosfolípides e proteínas de choque térmico (Hsp). O objetivo deste estudo foi investigar a presença de anticorpos contra fosfolípides, beta2-glicoproteína 1 (beta2-gp1), lipoproteína lipase (LPL) e Hsp em pacientes com AR e avaliar a associação entre estes anticorpos com a presença de aterosclerose subclínica de carótidas. Métodos: Anticorpos contra cardiolipina (aCL) IgG e IgM, beta2-gp1 IgG, IgM e IgA , Hsp 60 e Hsp 65 foram testados por ELISA em um grupo de 71 pacientes com AR comparado com 53 indívíduos controles não portadores de AR, de idade e sexo similar. Foram excluídos os pacientes com HAS, diabetes melitos e os fumantes em ambos os grupos. Níveis de lipoproteínas, parâmetros clínicos da AR, questionário de avaliação de saúde (HAQ), escore de atividade da doença (DAS) 28, velocidade de hemossedimentação (VHS) e proteína C reativa (PCR) foram avaliadas. A associação entre a presença dos anticorpos aCL, beta2-gp1, Hsp 60 e Hsp 65 com os parâmetros clínicos de atividade da doença, com a presença das placas de aterosclerose e com a medida da espessura íntimomedial (IMT) da carótida comum, usando ultra-som (US) modo B de alta resolução foram pesquisadas. Resultados: A idade média no grupo com AR foi 48,93 ± 12,31 vs. 45,37 ± 9,37 no grupo controle saudável (p = 0,20); 90,1% no grupo com AR eram do sexo feminino vs. 86,8% no grupo controle (p = 0,56); índice de massa corporal (IMC) foi 25,72 ± 4,57kg/m² no grupo com AR vs. 26,40 ± 4,52kg/m² no grupo controle (p = 0, 69); Os níveis de colesterol, LDL, triglicerídeos e a relação CT/HDL não foram diferentes quando comparamos os 2 grupos (p > 0,05). O nível de HDL foi maior no grupo com AR vs. grupo controle com 60,56 ± 14,40mg/dl e 54,52 ± 11,55 respectivamente (p = 0,05). A média da medida da IMT foi 0,721 ± 0,16 mm na AR e 0,667 ± 0,14mm no grupo controle, e a IMT dos pacientes com AR foi maior naqueles com idade acima dos 50 anos (P < 0,001). No grupo com AR, 14,1% dos pacientes tinham placas nas carótidas vs. 1,9% dos indivíduos saudáveis (p = 0,02) e no grupo com AR, as placas foram mais frequentes nos pacientes acima dos 50 anos (p = 0,004). No grupo AR, 5,6% tinham anticorpos aCL IgG vs. 3,8% no grupo controle (p > 0,05); 14,1% apresentavam aCL IgM vs. 7,5% (p > 0,05); 43,7% tinham anti-beta2-gp1 IgA vs. 40,8% no grupo controle (p > 0,05). A prevalência de anti-beta2-gp1 IgG e IgM e anti-LPL não foi diferente entre os pacientes com AR e o grupo controle ( p > 0,05). A presença dos anticorpos anti-Hsp 60 e 65 na AR e no grupo controle não foram diferentes (p > 0,05), mas os títulos de anticorpos contra Hsp 65 e beta2-gp1 IgM foram maiores no grupo com AR ( p = 0,007 e p = 0,03 respectivamente). Nós não encontramos associação entre a presença e os títulos dos anticorpos aCL IgG e IgM, beta2-gp1 IgG, IgM e IgA, LPL e Hsp 60 e 65 com a presença de placas nas carótidas ou com a medida da IMT (p > 0,05). Discussão: Este estudo confirma achados anteriores da maior prevalência de aterosclerose carotídea nos pacientes com AR e sua correlação com idade, colesterol e LDL. Embora tenha se encontrado uma tendência a maior presença de anticorpos nos pacientes com AR, não houve relação entre a presença da aterosclerose mais prevalente nos pacientes com AR, com a auto-imunidade dirigida contra cardiolipina, beta2-gp1 ou Hsp. / Purpose: Many questions remain unanswered about the causes of accelerated atherosclerosis in patients with inflammatory systemic diseases such as rheumatoid arthritis (RA). Some studies have suggested the role of autoimmunity besides inflammation in the pathogenesis of atherosclerosis in general population and have also discussed the possible association with antibodies directed to phospholipids and heat shock proteins (Hsp). The aim of this study was to investigate the presence of antibodies against phospholipids, beta2-glycoprotein1 (beta2-gp1), lipoprotein lipase (LPL) and Hsp in RA subjects and evaluate the association between these antibodies with the presence of subclinical carotid atherosclerosis. Methods: Tests to antibodies against cardiolipin (aCL) IgG and IgM, beta2-gp1 IgG, IgM and IgA ,Hsp 60 and Hsp 65 were done by ELISA test in a group of 71 RA subjects compared with 53 age and sex-matched non-RA subjects. Smoking, diabetic and hypertensive patients were excluded in both groups. The lipoprotein levels, clinical parameters of RA, Health Assessment Questionnaire (HAQ), Disease Activity Score (DAS) 28, Erythrocyte Sedimentation Rate (ESR) and C-Reactive Protein (CRP) were evaluated. The association between the presence of antibodies against cardiolipin, beta2-gp1 and Hsp 60 and 65 with the clinical parameters of disease activity in RA, and with the presence of plaques and mean intimo-medial thickness (IMT) of common carotid using high-resolution B-mode ultrasound were assessed. Results: Mean age in RA group was 48.93 ± 12.31 vs. 45.37 ± 9.37 in healthy control group (p = 0.20); 90.1% were women in RA group vs. 86.8% in healthy control (p = 0.56); body mass index (BMI) were 25.72 ± 4.57 in RA group vs. 26.40 ± 4.52 in healthy control (p = 0.69). The levels of cholesterol, LDL, triglycerides, CT/HDL didn t have difference between the two groups (p > 0.05). The HDL was higher in RA group vs. control group with 60.56 ± 14.40mg/dl and 54.52 ± 11.55 respectively (p = 0.05). The mean IMT was 0.721 ± 0.16mm in RA and 0.667 ± 0.14mm in control group, and the IMT was higher in patients older than 50 years among RA subjects (p < 0.001). In RA subjects, 14.1% had carotid plaques vs. 1.9% in healthy controls (p = 0.02). In RA group, the carotid plaques were more frequent in patients older than 50 years (p = 0.004). In RA group, 5.6% had antibodies against cardiolipin IgG vs. 3.8% in control group (p > 0.05); 14.1% in RA group had anti-cardiolipin IgM vs. 7.5% (p > 0.05); 43.7% had anti-beta2-gp1 IgA vs 40.8% in control group (p > 0.05). The presence of anti-beta2-gp1 IgG and IgM, and anti-LPL didn t have significant difference between the groups (p > 0.05). The prevalence of antibodies to Hsp 60 and Hsp 65 were similar in RA and in control group (p > 0.05), but the titers of antibodies against Hsp 65 and beta2-gp1 IgM were higher in RA group (p = 0.007 and p = 0.03 respectively). We didn t find relationship between antibodies against cardiolipin IgG and IgM, or beta2-gp1 IgG, IgM and IgA, LPL, Hsp 60 and 65 with mean IMT or plaque carotid (p > 0.05). Discussion: This study confirms the great prevalence of carotid atherosclerosis in RA subjects and its correlation with age, cholesterol and LDL. Although it was found a tendency to have more autoantibodies in RA subjects, there weren t any link between atherosclerosis in RA with autoimmunity against cardiolipin, beta2-gp 1, LPL or Hsp.

Como demonstrado em estudos prévios, quando injetada em indivíduos, a microemulsão lipídica rica em colesterol sem proteína (LDE) que mimetiza a composição da LDL adquire apoE no plasma e é captada por receptores de LDL. No presente estudo, a LDE marcada com colesterol-H3(CL) e oleato de colesterol-C14(OC) foi injetada em 20 pacientes com doença arterial coronária antes da cirurgia de revascularização miocárdica. Fragmentos de aorta, artéria radial, artéria torácica interna, veia safena e pericárdio descartados durante a cirurgia foram coletados e analisados para radioatividade juntos com amostras seriadas de plasma. A contagem radioativa de LDE-OC foi maior do que a de LDE-CL em todas amostras de plasma coletadas durante 24h, entretanto a captação de LDE-CL foi expressivamente maior do que a do OC em todos os fragmentos. A captação de LDE-CL pela aorta foi cinco vezes maior do que a de LDE-OC (p=0,0379), quatro vezes maior na artéria torácica interna (p=0,033), dez vezes maior na veia safena (p=0,006) e quatro vezes maior no pericárdio (p=0,010). Apenas na artéria radial a captação não obteve significância estatística (p=0,053). Os estudos in vitro de captação celular, bloqueio e das marcações imuno-histoquímicas confirmaram os achados in vivo. Concluindo, a expressiva captação vascular do CL comparada com à do OC sugere que o CL dissocia-se a partir das partículas da microemulsão e precipita-se nos vasos. Considerando a LDE como um modelo de microemulsão artificial para a LDL, os resultados sugerem que este tipo de deposição do CL na parede vascular pode constituir um novo mecanismo para a aterogênese. / As shown in previous studies, when injected into subjects, a protein-free cholesterol-rich microemulsion (LDE) that mimics LDL composition acquires apoE in the plasma and is taken-up by LDL receptors. In the current study, LDE labeled with H3-Cholesterol (FC) and C14-Cholesteryl Oleate(CO) was injected into 20 coronary artery disease patients 24h before myocardial revascularization surgery. Fragments of aortic, radial, internal thoracic arteries, safenous vein and pericardium discarded during surgery were collected and analyzed for radioactivity together with serial plasma samples. The radioactive counting of LDE-CO was greater than that of LDE-FC in all the plasma samples collected over 24h, but the uptake of LDE-FC was markedly greater than that of CO in all fragments. The uptake of LDE-FC by aorta was 5-fold greater than that of LDE-CO (p=0,0379), 4-fold greater in the internal thoracic artery (p=0,033), 10-fold greater in safenous vein (p=0,006) and 4-fold greater in pericardium (p=0,010). Only in the radial artery the uptake didn\'t attains statistical significance (p=0,053). The in vitro studies of cell uptake, blocking and immunohistochemistry marks confirm the in vivo finds. In conclusion, the remarkably greater vessel tissue uptake of FC compared with CO suggests that FC dissociate from the microemulsion particles and precipitate in the vessels. Considering LDE as an artificial microemulsion model for LDL, the results suggests that this type of FC deposition in the arterial wall, might constitute a novel atherogenic mechanism.

Estudamos 62 pacientes, que com teste ergométrico positivo, manifestaram angina estável e estavam sob controle farmacológico. Todos apresentavam cinecoronariografia mostrando obstrução >70% em pelo menos uma das principais artérias coronárias. Objetivamos avaliar parâmetros eletrocardiográficos e de pressão arterial, durante procedimento odontológico restaurador sob anestesia local com e sem vasoconstritor em presença de doença arterial coronária. As idades variaram de 39 a 80, média de 58,7±8,8 anos, sendo 51 (82,3%) homens. Trinta pacientes foram randomizados para receber anestesia local com solução de lidocaína a 2% com adrenalina 1:100.000 e os demais para lidocaína a 2% sem vasoconstritor. Todos os pacientes foram submetidos à monitorização ambulatorial da pressão arterial (MAPA) e eletrocardiografia dinâmica por 24 horas, iniciados 2 horas antes do procedimento odontológico. Consideramos 3 períodos de registro: (1) basal - os 60 minutos que antecederam ao procedimento odontológico; (2) procedimento - desde o início da anestesia até o final do procedimento odontológico restaurador; (3) subseqüente completar das 24 horas. A análise de variância com medidas repetidas mostrou que houve elevação significativa da pressão arterial sistólica e diastólica do período basal para o procedimento nos dois grupos estudados (aproximadamente 14mmHg e 5 a 7mmHg) respectivamente, quando analisados separadamente e quando confrontados não apresentaram diferença de comportamento entre si. A freqüência cardíaca não se alterou nos dois grupos estudados. Depressão do segmento ST >1mm ocorreu em 10 (17,9%) pacientes; todos os eventos ocorreram no mínimo 2 horas após o término do procedimento odontológico. Extra - sístoles supra-ventriculares e/ou extra-sístoles ventriculares em número maior do que 10/hora estiveram presentes em 17 (30,4%) pacientes durante as 24 horas e durante o período do procedimento em 7 (12,5%), sendo 4 (13,8%) do grupo que recebeu anestesia sem adrenalina e 3 (11,1%) do grupo que recebeu anestesia com adrenalina e o teste Exato de Fisher não mostrou diferença entre os grupos. Concluímos que não houve diferença em relação ao comportamento de pressão arterial, freqüência cardíaca, evidência de isquemia e arritmias entre os grupos. O uso associado de vasoconstritor mostrou-se, portanto, seguro dentro dos limites do estudo. / We enrolled 62 patients with positive exercise stress test who presented with stable angina and were receiving drug therapy. All had a coronary angiography screening showing >70% obstruction in at least one of the main coronary arteries. The study aimed to compare electrocardiographic and blood pressure parameters during restorative dentistry procedure under local anesthesia, both with and without vasoconstrictor, in the presence of coronary artery disease. Ages ranged from 39 to 80, (mean ± SD) 58.7±8.8 years, 51 (82.3%) of them were male. Thirty patients were randomly assigned to receive 2% lidocaine local anesthesia with 1:100,000 epinephrine, the others receiving 2% lidocaine without vasoconstrictor. All the patients underwent ambulatory blood pressure and 24-hour Holter monitoring, beginning two hours ahead of the dental procedure. Recording were made during (1) baseline - 60-minute period before dental procedure began; (2) procedure - from beginning of anesthesia until the end of the procedure; and (3) subsequent 24-hour period. Analysis of variance with repeat measures showed significant diastolic and systolic blood pressure increases from baseline to the period of the procedure, in the two study groups (approximately 14 mm Hg, and 5 to 7 mm Hg, respectively); both in a separate analysis and in a comparative analysis no significant difference between them could be confirmed. Heart rate did not change in neither of the two groups. ST-segment >1 mm depression was detected in 10 (17.9%) patients; all these events occurred at least two hours after the end of the dentistry procedure. Premature supraventricular systoles and/or premature ventricular systoles in a greater number than 10/hour were seen in 17 (30.4%) patients in the 24-hours period after the procedure; during the procedure they occurred in 7 (12.5%) patients, of whom 4 (13.8%) were in the group without, and 3 (11.1%) in the group with vasoconstrictor. The Fisher\'s exact test revealed no difference between the groups. We concluded that there was no difference of blood pressure, heart rate, evidence of ischemia or arrhythmia episodes between the groups. Thus, the associated use of vasoconstrictor proved to be safe within the limits of this study

Chan Fat-Yiu. / Thesis (M.Sc.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 85-93). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.4 / SUMMARY --- p.5 / ABBREVIATIONS --- p.9 / LIST OF TABLES --- p.11 / LIST OF FIGURES --- p.13 / Chapter CHAPTER I --- INTRODUCTION --- p.14 / Chapter 1.1 --- The Historical Aspects of C-Reaction Protein --- p.15 / Chapter 1.2 --- Biochemistry of CRP --- p.16 / Chapter 1.3 --- Physiology of CRP --- p.18 / Chapter 1.4 --- Current Clinical Applications of Serum CRP Assay --- p.19 / Chapter 1.5 --- Recent Findings of CRP --- p.21 / Chapter 1.5.1 --- Pathophysiology of atherosclerosis --- p.22 / Chapter 1.5.2 --- A nother atherogenic risk factor: hs- CRP --- p.26 / Chapter 1.5.3 --- Can hs-CRP replace other risk factors? --- p.30 / Chapter 1.5.4 --- Altering hs-CRP result in medication --- p.32 / Chapter 1.6 --- Methods of Measurement of CRP Concentration --- p.33 / Chapter 1.7 --- Analytical Considerations in the Measurement of hs-CRP --- p.34 / Chapter CHAPTER II --- OBJECTIVES AND SIGNIFICANCE --- p.36 / Chapter 2.1 --- Objectives --- p.37 / Chapter 2.2 --- Issues and Problems --- p.37 / Chapter 2.3 --- Significance and Value of this Study --- p.38 / Chapter CHAPTER III --- MA TERIALS AND METHODS I Setting up the serum hs-CRP assay on the Hitachi 911 Analyzer --- p.39 / Chapter 3.1 --- Materials --- p.40 / Chapter 3.1.1 --- Reagents from Roche Diagnostics --- p.40 / Chapter 3.1.2 --- Reagents for the Beckman Coulter Array ® Analyzer --- p.40 / Chapter 3.1.3 --- In-house reagents --- p.41 / Chapter 3.2. --- Apparatus and Equipment --- p.41 / Chapter 3.2.1 --- Hitachi 911 Analyzer --- p.41 / Chapter 3.2.2 --- Beckman Coulter Array ® 360 Analyzer --- p.42 / Chapter 3.3 --- The Tina-quant a C-Reactive Protein (Latex) Ultrasensitive Assay --- p.42 / Chapter 3.3.1 --- Priniciple of the Dual-Radius Enhanced Latex (DuREL´ёØ) technology --- p.42 / Chapter 3.3.2 --- Assessment of Analytical Performance --- p.45 / Chapter CHAPTER IV --- MA TERIALS AND METHODS II Serum hs-CRP in Chinese chronic-renal-failure patients with atherosclerotic vascular disease or cardiac valve calcification --- p.48 / Chapter 4.1 --- Patient Recruitment --- p.49 / Chapter 4.2. --- Blood Specimens --- p.49 / Chapter 4.3 --- Assay Methods --- p.50 / Chapter 4.3.1 --- hs-CRP --- p.50 / Chapter 4.3.2 --- TC --- p.50 / Chapter 4.3.3 --- TG --- p.51 / Chapter 4.3.4 --- HDL-C --- p.51 / Chapter 4.3.5 --- LDL-C --- p.52 / Chapter 4.3.6 --- Apo A-1 --- p.52 / Chapter 4.3.7 --- Apo B --- p.53 / Chapter 4.3.8 --- Lp(a) --- p.53 / Chapter 4.4 --- Ultrasound measurement of carotid artery inter-media thickness --- p.53 / Chapter 4.5 --- Statistical analysis --- p.54 / Chapter CHAPTER V --- RESUTLSI Setting up the serum hs-CRP assay on the Hitachi 911 Analyzer --- p.55 / Chapter 5.1 --- Imprecision --- p.56 / Chapter 5.2 --- Linearity --- p.56 / Chapter 5.3 --- Recovery --- p.56 / Chapter 5.4 --- Detection Limit --- p.57 / Chapter 5.5 --- Carry-over --- p.57 / Chapter CHAPTER VI --- RESULTS II Serum hs-CRP in Chinese chronic-renal-failure patients with atherosclerotic vascular disease or cardiac valve calcification --- p.63 / Chapter 6.1 --- Patient Recruitment --- p.64 / Chapter 6.2 --- Chinese chronic-renal-failure patients with AVD --- p.64 / Chapter 6.3 --- Chinese chronic-renal-failure patients with CVC --- p.65 / Chapter CHAPTER VII --- DISCUSSION I Performance of the serum hs-CRP assay on the Hitachi 911 Analyzer --- p.75 / Chapter 7.1 --- "Imprecision, Detection Limit, Linearity, and Recovery of hs-CRP Assay" --- p.76 / Chapter 7.1.1 --- Imprecision --- p.76 / Chapter 7.1.2 --- Detection Limit --- p.76 / Chapter 7.1.3 --- Linearity --- p.76 / Chapter 7.1.4 --- Recovery --- p.77 / Chapter 7.2 --- Overall Performance --- p.77 / Chapter CHAPTER VIII --- DISCUSSION II Serum hs-CRP in Chinese chronic-renal-failure patients with atherosclerotic vascular disease or cardiac valve calcification --- p.79 / Chapter 8.1 --- CAPD Patients --- p.80 / Chapter 8.2 --- Serum hs-CRP Concentration of AVD and CVC Patients --- p.81 / Chapter 8.3 --- Other risk factors in AVD and CVC Patients --- p.82 / Chapter 8.4 --- Conclusion --- p.83 / REFERENCES --- p.85

Objetivos: Muitas questões permanecem sobre as causas da aterosclerose acelerada nos pacientes com doenças inflamatórias sistêmicas como a artrite reumatóide (AR). Estudos na população geral sugeriram que além da inflamação existe uma participação patogênica da auto-imunidade na aterosclerose e discutem a possível associação dos anticorpos contra fosfolípides e proteínas de choque térmico (Hsp). O objetivo deste estudo foi investigar a presença de anticorpos contra fosfolípides, beta2-glicoproteína 1 (beta2-gp1), lipoproteína lipase (LPL) e Hsp em pacientes com AR e avaliar a associação entre estes anticorpos com a presença de aterosclerose subclínica de carótidas. Métodos: Anticorpos contra cardiolipina (aCL) IgG e IgM, beta2-gp1 IgG, IgM e IgA , Hsp 60 e Hsp 65 foram testados por ELISA em um grupo de 71 pacientes com AR comparado com 53 indívíduos controles não portadores de AR, de idade e sexo similar. Foram excluídos os pacientes com HAS, diabetes melitos e os fumantes em ambos os grupos. Níveis de lipoproteínas, parâmetros clínicos da AR, questionário de avaliação de saúde (HAQ), escore de atividade da doença (DAS) 28, velocidade de hemossedimentação (VHS) e proteína C reativa (PCR) foram avaliadas. A associação entre a presença dos anticorpos aCL, beta2-gp1, Hsp 60 e Hsp 65 com os parâmetros clínicos de atividade da doença, com a presença das placas de aterosclerose e com a medida da espessura íntimomedial (IMT) da carótida comum, usando ultra-som (US) modo B de alta resolução foram pesquisadas. Resultados: A idade média no grupo com AR foi 48,93 ± 12,31 vs. 45,37 ± 9,37 no grupo controle saudável (p = 0,20); 90,1% no grupo com AR eram do sexo feminino vs. 86,8% no grupo controle (p = 0,56); índice de massa corporal (IMC) foi 25,72 ± 4,57kg/m² no grupo com AR vs. 26,40 ± 4,52kg/m² no grupo controle (p = 0, 69); Os níveis de colesterol, LDL, triglicerídeos e a relação CT/HDL não foram diferentes quando comparamos os 2 grupos (p > 0,05). O nível de HDL foi maior no grupo com AR vs. grupo controle com 60,56 ± 14,40mg/dl e 54,52 ± 11,55 respectivamente (p = 0,05). A média da medida da IMT foi 0,721 ± 0,16 mm na AR e 0,667 ± 0,14mm no grupo controle, e a IMT dos pacientes com AR foi maior naqueles com idade acima dos 50 anos (P < 0,001). No grupo com AR, 14,1% dos pacientes tinham placas nas carótidas vs. 1,9% dos indivíduos saudáveis (p = 0,02) e no grupo com AR, as placas foram mais frequentes nos pacientes acima dos 50 anos (p = 0,004). No grupo AR, 5,6% tinham anticorpos aCL IgG vs. 3,8% no grupo controle (p > 0,05); 14,1% apresentavam aCL IgM vs. 7,5% (p > 0,05); 43,7% tinham anti-beta2-gp1 IgA vs. 40,8% no grupo controle (p > 0,05). A prevalência de anti-beta2-gp1 IgG e IgM e anti-LPL não foi diferente entre os pacientes com AR e o grupo controle ( p > 0,05). A presença dos anticorpos anti-Hsp 60 e 65 na AR e no grupo controle não foram diferentes (p > 0,05), mas os títulos de anticorpos contra Hsp 65 e beta2-gp1 IgM foram maiores no grupo com AR ( p = 0,007 e p = 0,03 respectivamente). Nós não encontramos associação entre a presença e os títulos dos anticorpos aCL IgG e IgM, beta2-gp1 IgG, IgM e IgA, LPL e Hsp 60 e 65 com a presença de placas nas carótidas ou com a medida da IMT (p > 0,05). Discussão: Este estudo confirma achados anteriores da maior prevalência de aterosclerose carotídea nos pacientes com AR e sua correlação com idade, colesterol e LDL. Embora tenha se encontrado uma tendência a maior presença de anticorpos nos pacientes com AR, não houve relação entre a presença da aterosclerose mais prevalente nos pacientes com AR, com a auto-imunidade dirigida contra cardiolipina, beta2-gp1 ou Hsp. / Purpose: Many questions remain unanswered about the causes of accelerated atherosclerosis in patients with inflammatory systemic diseases such as rheumatoid arthritis (RA). Some studies have suggested the role of autoimmunity besides inflammation in the pathogenesis of atherosclerosis in general population and have also discussed the possible association with antibodies directed to phospholipids and heat shock proteins (Hsp). The aim of this study was to investigate the presence of antibodies against phospholipids, beta2-glycoprotein1 (beta2-gp1), lipoprotein lipase (LPL) and Hsp in RA subjects and evaluate the association between these antibodies with the presence of subclinical carotid atherosclerosis. Methods: Tests to antibodies against cardiolipin (aCL) IgG and IgM, beta2-gp1 IgG, IgM and IgA ,Hsp 60 and Hsp 65 were done by ELISA test in a group of 71 RA subjects compared with 53 age and sex-matched non-RA subjects. Smoking, diabetic and hypertensive patients were excluded in both groups. The lipoprotein levels, clinical parameters of RA, Health Assessment Questionnaire (HAQ), Disease Activity Score (DAS) 28, Erythrocyte Sedimentation Rate (ESR) and C-Reactive Protein (CRP) were evaluated. The association between the presence of antibodies against cardiolipin, beta2-gp1 and Hsp 60 and 65 with the clinical parameters of disease activity in RA, and with the presence of plaques and mean intimo-medial thickness (IMT) of common carotid using high-resolution B-mode ultrasound were assessed. Results: Mean age in RA group was 48.93 ± 12.31 vs. 45.37 ± 9.37 in healthy control group (p = 0.20); 90.1% were women in RA group vs. 86.8% in healthy control (p = 0.56); body mass index (BMI) were 25.72 ± 4.57 in RA group vs. 26.40 ± 4.52 in healthy control (p = 0.69). The levels of cholesterol, LDL, triglycerides, CT/HDL didn t have difference between the two groups (p > 0.05). The HDL was higher in RA group vs. control group with 60.56 ± 14.40mg/dl and 54.52 ± 11.55 respectively (p = 0.05). The mean IMT was 0.721 ± 0.16mm in RA and 0.667 ± 0.14mm in control group, and the IMT was higher in patients older than 50 years among RA subjects (p < 0.001). In RA subjects, 14.1% had carotid plaques vs. 1.9% in healthy controls (p = 0.02). In RA group, the carotid plaques were more frequent in patients older than 50 years (p = 0.004). In RA group, 5.6% had antibodies against cardiolipin IgG vs. 3.8% in control group (p > 0.05); 14.1% in RA group had anti-cardiolipin IgM vs. 7.5% (p > 0.05); 43.7% had anti-beta2-gp1 IgA vs 40.8% in control group (p > 0.05). The presence of anti-beta2-gp1 IgG and IgM, and anti-LPL didn t have significant difference between the groups (p > 0.05). The prevalence of antibodies to Hsp 60 and Hsp 65 were similar in RA and in control group (p > 0.05), but the titers of antibodies against Hsp 65 and beta2-gp1 IgM were higher in RA group (p = 0.007 and p = 0.03 respectively). We didn t find relationship between antibodies against cardiolipin IgG and IgM, or beta2-gp1 IgG, IgM and IgA, LPL, Hsp 60 and 65 with mean IMT or plaque carotid (p > 0.05). Discussion: This study confirms the great prevalence of carotid atherosclerosis in RA subjects and its correlation with age, cholesterol and LDL. Although it was found a tendency to have more autoantibodies in RA subjects, there weren t any link between atherosclerosis in RA with autoimmunity against cardiolipin, beta2-gp 1, LPL or Hsp.

Dislipidemia é implicada no processo aterosclerótico do LES. A descrição de aLPL no LES associado a hipertrigliceridemia levou-nos a analisar esse anticorpo no contexto da inflamação envolvida na aterogênese. aLPL foi encontrado em 38% dos pacientes com LES com altos níveis de triglicérides. Correlação positiva significante foi observada entre aLPL e PCR, VHS, SLEDAI, anti-DNA, anti-cardiolipina e CH100 baixo. Análise de regressão múltipla confirmou a forte associação entre aLPL e PCR. Esses dados dão suporte à associação entre inflamação, resposta imune e dislipidemia, introduzindo o aLPL como um novo componente nos complexos eventos da aterogênese do LES / Dyslipidemia is implicated in the atherosclerosis process of SLE. The description of aLPL in SLE associated with hypertrigliceridemia prompted us to analyze this antibody in the context of the inflammation involved in the atherogenesis. aLPL was found in 38 por cento of SLE patients with high levels of triglycerides. Significant positive correlation was observed between aLPL and CRP, ESR, SLEDAI, anti-DNA, anti-cardiolipin and low CH100. Multiple regression analysis confirmed the strong association between aLPL and CRP. These data support the link between inflammation, immune response and dyslipidemia, introducing anti-LPL as new player in the complex events of atherogenesis in SLE