As repetições ricas em leucina (LRR) são domínios presentes em diversas famílias de proteínas com diferentes funções, sendo responsáveis pela formação de uma estrutura capaz de estabelecer interações protéicas. Em decorrência do projeto de caracterização de um segmento do cromossomo 17 de L. amazonensis, identificamos um gene que codifica uma proteína de 72 kDa, contendo em sua região central, seis motivos LRR. Genes ortólogos estão presentes nos genomas de L. major e L. braziliensis. Observamos que o gene LRR17 é regulado de forma distinta ao longo dos ciclos biológicos de L. braziliensis e L. amazonensis. A proteína LRR17 de L. braziliensis e secretada tanto nos estágios promastigota como amastigota. Identificamos também a secreção da proteína LRR17 em promastigotas de L. amazonensis. Obtivemos mutantes hiperexpressores da proteína LRR17 em L. braziliensis e L. amazonensis. As linhagens mutantes foram mais infectivas em infecções de macrófagos in vitro quando comparadas com a linhagem selvagem. A proteína LRR17 parece estar envolvida no processo de invasão do parasita em infecções in vitro e no estabelecimento da infecção da forma amastigota de Leishmania. / Leucine-rich repeats (LRRs) are versatile binding motifs found in a variety of proteins involved in protein-protein interactions. The LaLRR17 gene, identified initially in the L. amazonensis genome, encodes a protein with 6 LRR in its central region, that is secreted to the cytoplasm of L. amazonensis-infected macrophages. An orthologue to LaLRR17 was identified in L. braziliensis chromosome 17. LRR17 gene expression is regulated differentially during the life cycle of L. braziliensis and L. amazonensis. The LbLRR17 protein is secreted in L. braziliensis promastigotes and amastigotes. To characterize the function of the LRR17 protein we obtained transgenic parasite lines of L. amazonensis overexpressing the LaLRR17 gene and of L. braziliensis overexpressing the LbLRR17 gene. The mutants were more infective to macrophages in vitro when compared with the wild type strains, indicating that the LRR17 protein may interact with macrophage molecules, modulating the cellular response to increase parasite survival.

Leung Ka-Kit. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 129-131). / Abstracts in English and Chinese. / Abstract --- p.1 / Chapter CHAPTER 1. --- introduction --- p.14 / Chapter 1.1 --- Preamble --- p.14 / Chapter 1.2 --- Biological background --- p.14 / Chapter CHAPTER 2. --- previous related work --- p.18 / Chapter 2.1 --- Protein functionality analysis --- p.18 / Chapter 2.1.1 --- Analysis from primary structure --- p.18 / Chapter 2.1.2 --- Analysis from tertiary structure --- p.20 / Chapter 2.2 --- Secondary structure prediction --- p.21 / Chapter 2.3 --- Motivation - Challenges from protein complexity --- p.22 / Chapter CHAPTER 3. --- mathematical representations for protein properties and sequence alignment --- p.24 / Chapter 3.1 --- Secondary structure sequence model --- p.24 / Chapter 3.2 --- Substitution matrix --- p.26 / Chapter 3.3 --- Gap --- p.26 / Chapter 3.4 --- Similarity measurement --- p.27 / Chapter 3.5 --- Geometric Model for Protein --- p.28 / Chapter CHAPTER 4. --- overall system design --- p.30 / Chapter 4.1 --- System architecture and design --- p.30 / Chapter 4.2 --- System environment --- p.32 / Chapter 4.3 --- Experimental data --- p.32 / Chapter CHAPTER 5. --- adaptive dynamic programming (adp)- general global alignment consideration --- p.35 / Chapter 5.1 --- t-triangles cutting --- p.35 / Chapter 5.1.1 --- Theoretical time and memory requirements of ADP with z-triangles cutting --- p.43 / Chapter 5.1.1.1 --- Study of parameters affecting h in case 1 --- p.44 / Chapter 5.1.1.2 --- Study of parameters affecting h in case 2 --- p.45 / Chapter 5.1.2 --- Experimental results of ADP with z-triangles cutting --- p.46 / Chapter 5.2 --- Constructing the path matrix by expansion --- p.51 / Chapter 5.2.1 --- Time and memory requirements of EXPAND --- p.57 / Chapter 5.2.2 --- Experimental results and discussions --- p.58 / Chapter CHAPTER 6. --- adp - global alignment of sequences with consecutive repeated characters --- p.65 / Chapter 6.1 --- Estimation of similarity upper bound (Ba) --- p.65 / Chapter 6.1.1 --- Sequence composition (SC) consideration --- p.65 / Chapter 6.1.2 --- Implementation of SC --- p.67 / Chapter 6.1.3 --- Experimental results --- p.69 / Chapter 6.1.4 --- Overall trend of change of structures (OTCS) --- p.74 / Chapter 6.1.5 --- Uninformed search --- p.76 / Chapter 6.2 --- Short-cut --- p.80 / Chapter 6.2.1 --- Time and memory requirements --- p.86 / Chapter 6.2.2 --- Experimental results and discussions --- p.86 / Chapter CHAPTER 7. --- ga based topology discovery --- p.87 / Chapter 7.1 --- Chromosome encoding --- p.87 / Chapter 7.2 --- Non-sequential order penalty --- p.88 / Chapter 7.3 --- Fitness function --- p.88 / Chapter 7.4 --- Genetic operators --- p.88 / Chapter 7.4.1 --- Hop operator --- p.89 / Chapter 7.4.2 --- Inverse operator --- p.89 / Chapter 7.4.3 --- Shift operator --- p.90 / Chapter 7.4.4 --- Selection pressure --- p.90 / Chapter 7.5 --- Selection of progeny --- p.91 / Chapter 7.6 --- Implementation --- p.91 / Chapter 7.6.1 --- Size of population and generation --- p.91 / Chapter 7.6.2 --- Parallelization --- p.91 / Chapter 7.6.3 --- Crowding Handling --- p.92 / Chapter 7.6.4 --- Selection of progeny --- p.92 / Chapter 7.7 --- Results of alignment with GA exploration on topological order --- p.93 / Chapter CHAPTER 8. --- FILTERING OF FALSE POSITIVES --- p.103 / Chapter 8.1 --- Alignment Segments to Gap Ratio (ASGR) --- p.103 / Chapter 8.2 --- Tolerance --- p.104 / Chapter 8.3 --- Overall trend of change of structures (OTCS) --- p.104 / Chapter 8.4 --- Results and discussions --- p.105 / Chapter CHAPTER 9. --- SECONDARY STRUCTURE PREDICTION --- p.111 / Chapter 9.1 --- 3-STATE SECONDARY STRUCTURE PREDICTION IMPROVEMENT --- p.111 / Chapter 9.2 --- 8-state secondary structure prediction --- p.117 / Chapter 9.3 --- Iterative Subordinate Voting (IS V) --- p.117 / Chapter 9.4 --- ISV Results and discussion --- p.119 / Chapter CHAPTER 10. --- CONCLUSIONS --- p.123 / Chapter 10.1 --- Contributions --- p.123 / Chapter 10.2 --- Future Work --- p.126 / Chapter 10.2.1 --- Using database indexing --- p.126 / Chapter 10.2.2 --- 3-state secondary structure prediction improvement --- p.127 / appendix --- p.128 / Chapter ´Ø --- Interpretation on the dp一filter results --- p.128

The (Chilean) flat oyster, Ostrea chilensis, is native to New Zealand and the west coast of South America. It is a commercially important species in New Zealand because of its exquisite taste that attracts premium prices. This thesis describes the first isolation and partial charcterisation of an oyster haemolymph calcium-dependent carbohydrate-binding protein. This protein 'chiletin' was originally isolated from oyster haemolymph by binding to the agarose-galactan matrix of a Sepharose column. Chiletin was predominantly composed of a 24 kilodalton (kDa) band when examined with one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis under non-reducing conditions and a 12 kDa band with reduction of disulphide bonds. The N-terminal sequence of the 24 kDa band was determined to be 'IAGPGWEKYN'. This sequence was not homologous to any known protein. Examination of isolated chiletin with two-dimensional protein analysis gel electrophoresis revealed the presence of three (~12 kDa) subunits ranging in isoelectric point from 5.2 to 6.0. The 24 kDa protein was used to immunise rabbits and a separate antiserum was also raised in rabbits using a synthetic peptide (identical to that above) coupled to keyhole limpet haemocyanin. These antisera were used to confirm the size of the chiletin subunits with Western blots and to examine the elution of chiletin in oyster haemolymph with size exclusion chromatography in phosphate buffered saline (PBS) and 8 M urea. There were four or five different sized conformational aggregates of chiletin present in oyster haemolymph under physiological conditions (PBS). The use of 8 M urea produced two separate aggregates. A major characteristic of lectins is the ability to agglutinate sheep red blood cells and both whole oyster haemolymph and isolated chiletin had this property. Chiletin was identified by immunohistochemistry to be present in a number of tissues. Staining intensity was most consistent in the auricular myocardial cells, followed by the digestive gland epithelium. Chiletin was not induced in haemolymph in response to temperature (30??C) stress or injection of turpentine into the adductor muscle. There have been few immunological studies performed with O. chilensis. The results of the project contribute to what is known about comparative immunology. Greater understanding of how oysters respond to stress and deal with pathogens will ultimately be of benefit to the aquaculture industry.

Submitted by Vasti Diniz (vastijpa@hotmail.com) on 2017-09-08T11:51:16Z No. of bitstreams: 1 arquivototal.pdf: 7392216 bytes, checksum: dabe1bebbcfee7c351e5b74904f8fca2 (MD5) / Made available in DSpace on 2017-09-08T11:51:16Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 7392216 bytes, checksum: dabe1bebbcfee7c351e5b74904f8fca2 (MD5) Previous issue date: 2017-04-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Bothrops mattogrossensis snake was originally described in 1925 by Amaral, however, studies with this species are still scarce. The objective of this study was to evaluate and compare the biochemical and toxic effects of crude venom pools extracted from Bothrops mattogrossensis males (PB_BmatM) and females (PB_BmatF). The protein profile of each venom was characterized and the hemolytic, oxidant and coagulant effects were evaluated and compared between males and females. Chromatographic profiles and protein band patterns showed similarities between pools. Erythrocytes of types A and B present a higher percentage of hemolysis in the presence of both venoms and were more susceptible to hemolysis in the presence of fatty acids and calcium chloride, suggesting a possible action of phospholipases A2. During the hemolytic activity, visible hemagglutination was observed in the highest amounts of PB_BmatF and PB_BmatM evaluated. When compared to the control, the reduction in percentages of hemoglobins oxidized to methaemoglobin in the presence of 1000μg of the venoms were 30% (BmatM) and 35% (BmatF), whereas in 100μg BmatM it was 22.5%. PB_BmatM presented lower plasma coagulation time and both venoms presented dose-dependent activity inversely proportional to coagulation time. Therefore, the description and the comparative evaluation of the biochemical and toxinological effects triggered by the crude venoms of males and females of B. mattogrossensis provided additional information on the toxicity, in vitro, of these venoms. Still, these results open new questions and demand on the performance of the venom in front of the different blood types of the ABO system. / A serpente Bothrops mattogrossensis foi originalmente descrita em 1925 por Amaral, apesar disso, estudos com essa espécie ainda são escassos. O objetivo desse estudo foi avaliar e comparar os efeitos bioquímicos e tóxicos de pools de peçonhas brutas extraídas de machos (PB_BmatM) e fêmeas (PB_BmatF) de Bothrops mattogrossensis. O perfil proteico de cada peçonha foi caraterizado e os efeitos hemolítico, oxidante e coagulante foram avaliados e comparados entre machos e fêmeas. Os perfis cromatográficos e os padrões de bandas proteicas mostraram semelhanças entre os pools. Verificou-se que, eritrócitos dos tipos A e B apresentam maior percentual de hemólise na presença de ambos as peçonhas e foram mais susceptíveis à hemólise na presença de ácidos graxos e cloreto de cálcio, sugerindo uma possível ação das fosfolipases A2. Durante a realização da atividade hemolítica observou-se hemaglutinação visível nas maiores quantidades de PB_BmatF e PB_BmatM avaliadas. Quando comparado ao controle, a redução nos percentuais de hemoglobinas oxidadas à metahemoglobina na presença de 1000μg das peçonhas foram de 30% (BmatM) e 35% (BmatF), enquanto que em 100μg de BmatM foi de 22,5%. A PB_BmatM apresentou menor tempo de coagulação e ambas as peçonhas apresentaram atividade dose dependente. Portanto, a descrição e a avaliação comparativa dos efeitos bioquímicos e toxinológicos desencadeados pelas peçonhas brutas de machos e fêmeas de B. mattogrossensis forneceram informações adicionais sobre à toxicidade, in vitro, dessas peçonhas. Ainda, esses resultados poderão abrir novas indagações e buscas sobre sua atuação frente aos diferentes tipos sanguíneos do sistema ABO.

As repetições ricas em leucina (LRR) são domínios presentes em diversas famílias de proteínas com diferentes funções, sendo responsáveis pela formação de uma estrutura capaz de estabelecer interações protéicas. Em decorrência do projeto de caracterização de um segmento do cromossomo 17 de L. amazonensis, identificamos um gene que codifica uma proteína de 72 kDa, contendo em sua região central, seis motivos LRR. Genes ortólogos estão presentes nos genomas de L. major e L. braziliensis. Observamos que o gene LRR17 é regulado de forma distinta ao longo dos ciclos biológicos de L. braziliensis e L. amazonensis. A proteína LRR17 de L. braziliensis e secretada tanto nos estágios promastigota como amastigota. Identificamos também a secreção da proteína LRR17 em promastigotas de L. amazonensis. Obtivemos mutantes hiperexpressores da proteína LRR17 em L. braziliensis e L. amazonensis. As linhagens mutantes foram mais infectivas em infecções de macrófagos in vitro quando comparadas com a linhagem selvagem. A proteína LRR17 parece estar envolvida no processo de invasão do parasita em infecções in vitro e no estabelecimento da infecção da forma amastigota de Leishmania. / Leucine-rich repeats (LRRs) are versatile binding motifs found in a variety of proteins involved in protein-protein interactions. The LaLRR17 gene, identified initially in the L. amazonensis genome, encodes a protein with 6 LRR in its central region, that is secreted to the cytoplasm of L. amazonensis-infected macrophages. An orthologue to LaLRR17 was identified in L. braziliensis chromosome 17. LRR17 gene expression is regulated differentially during the life cycle of L. braziliensis and L. amazonensis. The LbLRR17 protein is secreted in L. braziliensis promastigotes and amastigotes. To characterize the function of the LRR17 protein we obtained transgenic parasite lines of L. amazonensis overexpressing the LaLRR17 gene and of L. braziliensis overexpressing the LbLRR17 gene. The mutants were more infective to macrophages in vitro when compared with the wild type strains, indicating that the LRR17 protein may interact with macrophage molecules, modulating the cellular response to increase parasite survival.

Ces travaux de recherche s’intéressent à la conception de nouvelles méthodologies analytiques destinées à mesurer le bénéfice d’une transplantation hépatique ou bien encore à l’évaluation de nouvelles voies thérapeutiques à l’essai pour des patients atteints de la polyneuropathie amyloïde familiale à transthyrétine (TTR). Cette maladie rare se caractérise par une déstabilisation structurale du tétramère de TTR qui aboutit à l’agrégation de fibrille dans les tissus du système nerveux autonome, au niveau des nerfs périphériques et autour de certains organes dont le cœur. Dans le cadre d’une collaboration entre hospitalo-universitaires, chimistes analystes, électrochimistes, physico-chimistes et technologues, nous nous sommes attachés à développer des séparations en électrophorèse capillaire couplée avec une détection optique de peptides natif et muté qui sont directement associés à une variente de cette maladie rare. La difficulté première de cette recherche concerne le choix même de ces biomarqueurs qui s’est finalement révélé pertinent grâce de la réalisation de cartes peptidiques à partir du sérum. Ensuite deux voies ont été explorées : une séparation électrocinétique avec une détection spectrométrique d’absorbance dans l’ultra-violet et l’autre nécessitant le marquage préalable de peptides par des molécules fluorophores ou fluorogènes pour ensuite faire une séparation en électrophorèse couplée LIF (laser induced fluorescence). Dans les deux cas le critère principal de séparation, la résolution, autorise une quantification et surtout les validations analytiques associées montrent une réelle robustesse des méthodologies développées. L’autre signe encourageant pour la transposition des ces méthodes à l’analyse de prélèvements issus de patients, concerne la limite de quantification qui est inférieure à celle couramment mesuré dans le sérum. La spectrométrie de masse, moyen d’investigation physico-chimique puissant à permis de suivre et de comprendre d’un point vue plus fondamental le produit des réactions de chimie organique de dérivation des peptides par trois marqueurs fluorescents : le TAMRA-SE, le NDA et le FQ. La possibilité de proposer un outil d’analyse miniaturisé et simple d’utilisation pour le monde hospitalier a également été étudiée. Un poste d’analyse sur puce microfluidique permettant l’analyse quantitative et qualitative a été installé pour permettre la réalisation de premiers essais expérimentaux de séparations électrocinétiques sur puce microfluidique. Ces travaux jettent les bases d’une nouvelle voie analytique pour séparer et quantifier les différents biomarqueurs caractéristiques de la polyneuropathie amyloïde familliale à TTR. / The purpose of our work was the development of new analytical methodologies to measure the benefit of liver transplantation and also the evaluation of new therapeutic approaches under testing on patients with Transtyretin (TTR) familial amyloid polyneuropathy. This rare disease is characterized by a structural destabilization of TTR tetramer leading to it’s aggregation into amyloïd fibrils that accumulate in the tissues of the autonomous nervous system, peripheral nerves and around certain organs, including the heart. As part of a collaboration between university, hospital, analytical chemists, electrochemist, physical chemists and technologists, we are committed to develop separations in capillary electrophoresis coupled with optical detection of native and mutated peptides that are directly associated with a variant of this rare disease. The first challenge of this research is the choice of these biomarkers that ultimately proved relevant with the realization of peptide maps from the serum. Then two approaches have been explored: electrokinetic separation with absorbance spectrometric detection in the ultraviolet and the other requiring the prior labeling peptides with fluorescent molecules and then to a separation on electrophoresis coupled with LIF (Laser induced fluorescence). In both cases the main criterion of separation, resolution, allows quantification and especially analytical validations show actual strength associated methodologies developed. Another encouraging sign for the transposition of these methods to the analysis of samples from patients regarding the quantification limit is lower than commonly measured in serum. Mass spectrometry, using physico-chemical investigation powerful allowed to follow and understand a more fundamental viewpoint the product of organic chemistry reactions bypass peptides by three fluorescent dyes: TAMRA-SE, the NDA and FQ. The ability to provide a miniaturized analysis and easy to use tool for the hospital environment was also studied. A post analysis on microfluidic chip for quantitative and qualitative analysis was installed to allow the realization of the first experimental tests of electrokinetic separations on microfluidic chip. These studies lay the foundation for a new analytical way to separate and quantify the different characteristics biomarkers family TTR amyloid polyneuropathy.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / The Brazilian spotted fever is an infectious disease transmitted by ticks to humans. The occurrence of Rickettsia rickettsii agent has been reported in Brazil since 1920 and is considered the main bacteria involved in Brazilian Spotted Fever. Since 2000, four other current species have been identified in the country, as follows: Rickettsia riphicephali, Rickettsia amblyommi, Rickettsia parkeri and Rickettsia felis. R. parkeri was first isolated in Amblyomma maculatum on the Gulf Coast of the United States in 1937, but until 2004 was considered as non-pathogenic agent, when was the first recognized case of spotted fever in humans caused by this species. Recently a new human rickettsial infection was reported to cause disease in São Paulo, being called Rickettsia parkeri Strain Atlantic Forest. This study aimed to detect and identify proteins with potential to stimulate the immune system of the host of this new strain described. Therefore, we performed total protein extraction R. parkeri Strain Atlantic Forest from infected VERO cell samples. The extracted proteins were fractionated by electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate (SDS-PAGE). The proteins were transferred to nitrocellulose membranes by semi-dry electrotransfer system and subjected to Western blot. Subsequently, the membrane was incubated with domestic rabbit hyperimmune serum against R. parkeri Strain Atlantic Forest (primary antibodies) followed by incubation with anti-rabbit IgG peroxidase-conjugated antibody (secondary antibody) to detect the primary antibodies bound to the proteins . Obtaining the hyperimmune rabbit serum was performed by experimental infection of R. parkeri Strain Atlantic Forest in domestic rabbit, intraperitoneally. After incubation, the disclosure of immunoreactive proteins was performed using a chromogenic substrate. 2 immunoreactants were detected proteins with more than 78 kDa (200 e 130 kDa) and 5 with less than 78 kDa. By comparing existing proteomic maps and the molecular weight of these proteins showed that, it is suggested that rOmpA (200 kDa) and rOmpB (130 kDa) are among the proteins detected and the remaining proteins found are members of the family of surface antigens cell (Sca - Surface cell antigen). However, one can not say that these proteins represent only reactive immunity to R. parkeri Strain Atlantic Forest, or that are homologous to other species. However, studies using the Modern Liquid Chromatography technique associated with mass spectrometry (LC/MS/MS) must be conducted for protein characterization. Preliminary results obtained in this study allowed further elucidation of protein profile of this agent, providing subsidies for the development of highly sensitive and specific diagnoses, as well as studies allow for the realization of a vaccine, as an alternative for the control and prevention of Brazilian Spotted Fever. / A Febre Maculosa Brasileira é uma doença infecciosa, transmitida por carrapatos ao homem. A ocorrência do agente Rickettsia rickettsii tem sido relatada no Brasil desde 1920, sendo considerada a principal bactéria envolvida na Febre Maculosa Brasileira. Desde 2000, outras quatros espécies circulantes foram identificadas no país, sendo: Rickettsia riphicephali, Rickettsia amblyommi, Rickettsia parkeri e Rickettsia felis. R. parkeri foi isolada pela primeira vez em Amblyomma maculatum, na Costa do Golfo dos Estados Unidos da América em 1937, porém até 2004 era considerada como agente não patogênico, quando então houve o primeiro caso reconhecido de Febre Maculosa em humanos ocasionado por essa espécie. Uma nova riquetsiose humana foi descrita como causadora da doença no Estado de São Paulo, sendo denominada de Rickettsia parkeri cepa Mata Atlântica. O presente trabalho teve como objetivo detectar e identificar proteínas com potencial de estimular o sistema imune do hospedeiro desta nova cepa descrita. Para tanto, foi realizado a extração proteica total de R. parkeri cepa Mata Atlântica a partir de amostras de células VERO infectadas. As proteínas extraídas foram fracionadas por eletroforese em gel de poliacrilamida na presença de dodecil-sulfato de sódio (SDS-PAGE). Estas proteínas foram transferidas para membranas de nitrocelulose através do sistema semi-seco de eletrotransferência e submetidas à técnica de Western blot. Posteriormente, a membrana foi incubada com soro hiperimune de coelho doméstico contra R. parkeri cepa Mata Atlântica (anticorpos primários), seguido de incubação com anticorpo anti-IgG de coelho conjugado com peroxidase (anticorpo secundário), para detectar os anticorpos primários ligados às proteínas. A obtenção do soro de coelho hiperimune foi realizada através da infecção experimental de R. parkeri cepa Mata Atlântica em um coelho doméstico, via intraperitoneal. A revelação das proteínas imunorreativas foi efetuada através de substrato cromogênico. Foram detectadas 2 proteínas imunorreagentes com mais de 78 kDa (200 e 130 kDa ) e 5 com menos de 78 kDa. Através da comparação de mapas proteômicos existentes e pelo peso molecular que estas proteínas apresentaram, sugere-se que rOmpA (200 kDa) e rOmpB (130 kDa) estejam entre as proteínas detectadas e as demais proteínas encontradas, sejam membros da família de antígenos de superfície celular (Sca Surface cella ntigen). No entanto, não se pode afirmar que estas proteínas apresentam imunidade reativa única para R. parkeri cepa Mata Atlântica, ou se são homólogas a outras espécies de riquetsias. Contudo, estudos empregando a técnica de Cromatografia Líquida Moderna associada a Espectrometria de Massas (LC/MS/MS) deverão ser conduzidos para a caracterização proteica. Os resultados preliminares obtidos neste estudo permitiram maior elucidação do perfil proteico deste agente, fornecendo assim subsídios para o desenvolvimento de diagnósticos altamente sensíveis e específicos, bem como permitir estudos para a realização de uma vacina, como alternativa para o controle e prevenção da Febre Maculosa Brasileira.

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

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

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