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Atherosclerosis is considered a chronic inflammatory disease of the arterial vessel wall which is not only modulated by innate and adaptive immune responses but also by factors of the blood coagulation system. In general hypercoagulability seems to increase the development and progression of experimental atherosclerosis in mice on an atherogenic background. In addition, the great majority of coagulation proteins including coagulation factor XII (FXII) have been detected in early and advanced human atherosclerotic lesions supporting the cross-link between the coagulation system and atherosclerosis. Moreover, FXII has been detected in close proximity to macrophages, foam cells and smooth muscle cells in these lesions and has been demonstrated to be functionally active in human plaques. Although these data indicate that factor XII may play a role in atherogenesis a direct contribution of FXII to atherogenesis has not been addressed experimentally to date. Furthermore, clinical studies examining the function of FXII in vascular disease have yielded conflicting results. Hence, in order to investigate the function of coagulation factor XII in atherosclerosis apolipoprotein E and FXII-deficient (F12\(^{-/-}\) apoE\(^{-/-}\)) mice were employed. Compared to F12\(^{+/+}\)apoE\(^{-/-}\) controls, atherosclerotic lesion formation was reduced in F12\(^{-/-}\)apoE\(^{-/-}\) mice, associated with diminished systemic T-cell activation and Th1-cell polarization after 12 weeks of high fat diet. Moreover, a significant decrease in plasma levels of complement factor C5a was evidenced in F12\(^{-/-}\)apoE\(^{-/-}\) mice. Interestingly, C5a increased the production of interleukin-12 (IL-12) in dendritic cells (DCs) and enhanced their capacity to trigger antigen-specific interferon-gamma (IFNγ) production in OTII CD4\(^+\) T cells in vitro. Importantly, a reduction in frequencies of IL-12 expressing splenic DCs from atherosclerotic F12\(^{-/-}\)apoE\(^{-/-}\) versus F12\(^{+/+}\)apoE\(^{-/-}\) mice was observed in vivo, accompanied by a diminished splenic Il12 transcript expression and significantly reduced IL-12 serum levels. Consequently, these data reveal FXII to play an important role in atherosclerotic lesion formation and to promote DC-induced and systemic IL 12 expression as well as pro-inflammatory T-cell responses likely at least in part via the activation of the complement system. / Die Arteriosklerose wird als eine chronisch entzündliche Erkrankung der arteriellen Gefäßwand angesehen, welche nicht nur durch Antworten des angeborenen und erworbenen Immunsystems, sondern auch durch Faktoren des Blutgerinnungssystems beeinflusst wird. Aus Tierstudien weiß man, dass Hyperkoagulabilität im Allgemeinen die Entwicklung und das Fortschreiten der Arteriosklerose in Mäusen mit atherogenem Hintergrund steigert. Ferner wurde die Mehrheit der Blutgerinnungsproteine inklusive des Koagulationsfaktors XII (FXII) in frühen und fortgeschrittenen arteriosklerotischen Läsionen in Menschen nachgewiesen, was eine mögliche Verbindung zwischen Koagulationssystem und Arteriosklerose untermauert. Darüber hinaus wurde der FXII in diesen Läsionen im unmittelbaren Umfeld von Makrophagen, Schaumzellen und glatten Muskelzellen nachgewiesen und dessen funktionelle Aktivität in humanen Plaques aufgezeigt. Obwohl diese Daten eine Funktion von FXII in der Arteriosklerose nahelegen, wurde eine direkte Beteiligung von FXII an der Atherogenese bislang experimentell noch nicht untersucht. Ferner erzielten klinische Studien, welche die Funktion von FXII in Gefäßerkrankungen untersuchten, widersprüchliche Resultate. Um die Funktion des Koagulationsfaktors XII in der Atherosklerose zu untersuchen, wurden daher Apolipoprotein E und FXII-defiziente (F12\(^{-/-}\)apoE\(^{-/-}\)) Mäuse eingesetzt. Die Ausbildung arteriosklerotischer Läsionen war in F12\(^{+/+}\)apoE\(^{-/-}\) Mäusen im Vergleich zu F12\(^{+/+}\)apoE\(^{-/-}\) Kontrollen reduziert. Dies ging mit einer verringerten systemischen T-Zell Aktivierung und Th1-Zell Polarisierung nach 12-wöchiger atherogener Diät einher. Ferner konnte eine signifikante Reduktion der C5a-Plasmaspiegel in F12\(^{-/-}\)apoE\(^{-/-}\) Mäusen nachgewiesen werden. Interessanterweise konnte C5a die Interleukin-12 (IL 12) Produktion in dendritischen Zellen (DCs) steigern, sowie deren Fähigkeit erhöhen, eine antigen-spezifische Interferon-gamma (IFNγ) Produktion in vitro, in OTII CD4\(^+\) T-Zellen zu induzieren. Insbesondere konnte in vivo eine Reduktion IL-12-exprimierender DCs in Milzen arteriosklerotischer F12\(^{-/-}\)apoE\(^{-/-}\) im Vergleich zu F12\(^{+/+}\)apoE\(^{-/-}\) Mäusen beobachtet werden, welche mit einer verminderten Expression an Il12 Transkripten in der Milz und signifikant reduzierten IL-12 Serumspiegeln einherging. Zusammenfassend zeigen diese Daten, dass FXII eine wichtige Rolle in der Bildung arteriosklerotischer Läsionen spielt und die DC-vermittelte und systemische IL-12 Expression, sowie pro-inflammatorische T-Zell Antworten, vermutlich teilweise über eine Aktivierung des Komplementsystems, fördert.

Protamin antagonisiert die antikoagulierende Wirkung von Heparin. Nach intravenöser Protaminapplikation treten als häufige unerwünschte Wirkungen ein systemischer Blutdruckabfall, Herzfrequenzabfall sowie eine Erhöhung des pulmonalarteriellen Widerstandes auf. Die Protamin-assoziierten Nebenwirkungen sind zum Teil lebensbedrohlich. Der ihnen zugrunde liegende Mechanismus wurde in der vorliegenden Arbeit auf Zellkultur- und Gesamttierebene analysiert sowie mögliche Therapieoptionen aufgezeigt. Heparin-Protamin-Komplexe aktivieren auf Endothelzellen den Blutgerinnungsfaktor XII. Aktiver Faktor XII startet über sein Substrat Plasmakallikrein die Freisetzung des Peptidhormons Bradykinin aus hochmolekularem Kininogen. Funktions-inhibierende Antikörper oder pharmakologische Inhibitoren von Plasmakallikrein oder Faktor XII blockierten die Heparin-Protamin induzierte Bradykininbildung auf Zellen. Stickstoffmonoxid-spezifische Fluorophore zeigten, dass Bradykinin-Bindung an Kinin B2 Rezeptoren die endotheliale Stickstoffmonoxid-Synthase aktiviert. B2 Rezeptorantagonisten blockierten die Heparin-Protamin induzierte Stickstoff-monoxidbildung. Die intravenöse Infusion von Protamin in heparinisierte Wildtypmäuse senkte den systemischen Blutdruck und die Herzfrequenz. Im Gegensatz dazu waren Faktor XII und B2 Rezeptor Gen-defiziente Mäuse oder Tiere, die Faktor XII Inhibitoren oder B2 Rezeptorantagonisten infundiert bekamen, vor Heparin-Protamin-Effekten geschützt. Mit dieser Arbeit konnte gezeigt werden, dass Heparin-Protamin-Komplikationen durch eine Faktor XII-getriebene Bradykininbildung verursacht werden. Eine Blockade der Bradykininbildung oder -wirkung eröffnet eventuell eine Möglichkeit, die Heparin-Protamin-Nebenwirkungen auch beim Patienten zu therapieren. / Protamine is the antidot for anticoagulant effects of heparin. Intravenous application of protamine may cause systemic hypotension, heart rate decrease and increase in pulmonary pressure. Adverse effects associated with protamine are common and potentially life-threatening. This work characterizes the pathomechanism of heparin-protamine-effects in cell culture and in transgenic mice and suggests therapeutic options to block the adverse effects. Heparin-protamine-complexes activate blood coagulation factor XII on endothelial cells. Active factor XII initiates the generation of the vasoactive peptide hormone bradykinin from high molecular weight kininogen by plasmakallikrein action. Antibodies or low molecular weight inhibitors that interfere with factor XII or plasmakallikrein activity blocked heparin-protamine-induced bradykinin effects on cells. Nitric oxide-specific dyes revealed that bradykinin-binding to kinin B2 receptors activates the endothelial nitric oxide synthase. B2 receptor antagonists interfered with heparin/protamine-driven nitric oxide generation. Intravenous application of protamine into heparinized wildtype mice reduced systemic blood pressure and heart rate. Factor XII or B2 receptor deficient mice were protected from heparin-protamine-effects. Consistently, pharmacological targeting of B2 receptors or factor XII inhibited protamine associated cardiovascular effects in heparinized wildtype mice. Together, this work demonstrates that heparin-protamine-adverse effects are due to factor XII activation that culminates in the generation of bradykinin. Inhibition of bradykinin formation or signaling may offer novel strategies to block adverse heparin-protamin-effects in patients.

En filmanalys som fokuserar på att studera framställningen omkring den svenske kungen Karl XII. Studien är omfattad att analysera olika filmer med olika produktionsländer för att finna hur olika filmskapare och länder har valt att presentera den svenske kungen. Studien fokuserar på att undersöka den bild som ges av Karl XII i olika filmer för att finna hur historien omkring Karl XII har brukats.

SUMMARY Mast cell activation in allergic and inflammatory disease causes increased vascular permeability and edema. This thesis identifies a paracrine mechanism, by which heparin released from intracellular granules, is involved in mast cell-evoked alteration of endothelial barrier function in vivo. Negatively charged heparin initiated factor XII-driven contact activation. Activated factor XII triggered the formation of the inflammatory mediator bradykinin in plasma. Congenital deficiency and pharmacological targeting of factor XII and kinin B2 receptor provided protection from mast cell-heparin-induced leukocyte-endothelial adhesion and hypotension in rats and mice. Intravital laser scanning microscopy and tracer measurements showed that heparin increased leakage with fluid extravasation in skin microvessels in mice. Deficiency in factor XII or kinin B2 receptor conferred resistance to heparin-induced skin edema and largely protected mice from endothelial barrier dysfunction, caused by allergen-induced mast cell activation and anaphylactic reactions. In contrast, heparin and mast cell activation caused excessive edema formation in mice, deficient in the major inhibitor of factor XII, C1 esterase inhibitor. Hereditary angioedema patients, lacking C1 esterase inhibitor, suffered from allergeninduced edema. The data indicate that mast cell-heparin-initiated bradykinin formation plays a fundamental role in defective barrier function of pathological mast cell-mediated inflammation, hypotension and edema formation. / ZUSAMMENFASSUNG Aktivierte Mastzellen sind bei Allergien und Entzündungskrankheiten an der Ödembildung beteiligt. Diese Arbeit zeigt, wie von aktivierten Mastzellen freigesetztes Heparin die Gefäßpermeabilität erhöht. Mastzell-Heparin aktiviert im Plasma den Blutgerinnungsfaktor XII. Aktiver Faktor XII startet die Bildung des Entzündungsmediators Bradykinin durch Plasmakallikrein-vermittelte Spaltung von hochmolekularem Kininogen. Bradykinin führt zur Ödembildung und Vasodilatation. Bei Ratten und Mäusen blockieren Faktor XII- oder Kinin B2 Rezeptor-Inhibitoren die Heparin-induzierte und Bradykinin-vermittelte Leukozyten-Endothel Adhäsion und den arteriellen Blutdruckabfall. Um den Heparin-vermittelten Flüssigkeitsaustritt aus Mikrogefäßen in der Haut von Mäusen zu analysieren, wurde eine intravitale konfokale Mikroskopietechnik etabliert. Genetische Inaktivierung oder pharmakologische Blockierung von Faktor XII oder Kinin B2 Rezeptoren hemmen den Heparinvermittelten Flüssigkeitsaustritt. Sowohl in systemischen als auch in cutanen passiven Anaphylaxiemodellen waren Faktor XII- und Kinin B2 Rezeptor-defiziente Mäuse vor Allergen-induzierten Ödemen und anaphylaktischen Reaktionen geschützt. Im Gegensatz dazu führte eine Allergenexposition zu einer überschiessenden Ödembildung in C1 Esterase Inhibitor-defizienten Mäusen, bei denen das Gen für den wichtigsten Inhibitor von Faktor XII inaktiviert wurde. Bei Hereditären Angioödem- Patienten, denen funktionaler C1 Esterase Inhibitor fehlt, lösen allergischen Reaktionen Ödemattacken aus. Zusammenfassend zeigen diese Daten erstmals, dass die durch Heparin gestartet Bradykinin-Bildung, eine bedeutende Rolle bei der fehlerhaften Barrierenfunktion der pathologischen Mastzell-vermittelten Entzündungen, Blutdruckabfall und Ödembildung spielt.

Essen, Universiẗat, Diss., 2002.

Submitted by ELZA SILVA CARDOSO SOFFIATTI null (elza-cardoso@hotmail.com) on 2016-11-03T12:31:24Z No. of bitstreams: 1 Pio XII e as origens do Concílio Vaticano II_Elza Soffiatti.pdf: 1743380 bytes, checksum: 045c0b5d36a7e18701516b3618e49f4a (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-11-10T12:35:55Z (GMT) No. of bitstreams: 1 soffiati_esc_dr_fran.pdf: 1743380 bytes, checksum: 045c0b5d36a7e18701516b3618e49f4a (MD5) / Made available in DSpace on 2016-11-10T12:35:55Z (GMT). No. of bitstreams: 1 soffiati_esc_dr_fran.pdf: 1743380 bytes, checksum: 045c0b5d36a7e18701516b3618e49f4a (MD5) Previous issue date: 2016-10-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Esta tese apresenta o pensamento social católico da segunda metade do século XX, conforme o discurso do Papa Pio XII (1939-1958) e do Concílio Vaticano II (1962-1965). O argumento central é que a renovação trazida pelo Concílio Vaticano II, as bases eclesiais e as práticas religiosas católicas após a sua promulgação pelo papa Paulo VI já estavam em grande medida expressas nas proposições apresentadas pelo papa Pio XII nos 19 anos do seu pontificado. Tomando como aporte teórico a história das religiões, analisamos comparativamente os documentos produzidos pelo papa Pio XII (SDR – 1) e pelo Vaticano II; neste, principalmente, as Constituições Lumen Gentium e Gaudium et Spes (SDR – 2), com vistas a apreender a gênese das ideias religiosas, suas origens e desenvolvimento, as crenças derivadas e sua incidência nas práticas sociais das sociedades dos pós-guerra. No primeiro capítulo, discutimos as condições de enunciação do discurso religioso e suas interfaces com outras identidades e matizes discursivos, bem como as relações entre teologia e história como saber religioso e instrumento hermenêutico para o objeto de investigação. No capítulo seguinte, mostramos as diversas fórmulas adotadas pelo papa Pio XII para o anúncio do discurso católico, bem como a sua incidência sobre a natureza das relações sociais, internacionais, econômicas e religiosas, ambientadas no contexto do seu pontificado: durante e após a Segunda Guerra Mundial. No capítulo final, apontamos as teses de Pio XII em relação às teses conciliares do Vaticano II, destacando a compreensão em relação ao mundo de então, a proposta de uma ética social de matriz religiosa e cristã, ambas as iniciativas constituintes de um projeto de sociedade propugnado pelo catolicismo, e a ocupação por ele de um novo topoi enunciativo, que buscava acomodação na relação entre as esferas pública e privada, a partir do esforço de construção de referentes comunicacionais que facultassem o diálogo com o mundo contemporâneo.

Le facteur XII (FXII) est une sérine protéase de 80 kDa produite et sécrétée par le foie qui initie la voie intrinsèque de la coagulation. Diverses études ont montré un rôle délétère du FXII dans des pathologies cérébrales telles que l'accident vasulaie cérébral, la maladie d'Alzheimer et la slérose en plaques. Néanmoins, l'impact direct du FXII sur le devenir des cellules neuronales reste inconnu. De plus, l'expression du FXII dans le système nerveux central (SNC) n'a pas encore été étudiée. Le premier objectif de nos travaux a été d'étudier le rôle du FXII dans l'apoptose neuronale puis dans un second temps d'étudier l'expession du FXII dans le SNC. Dans notre étude, nous avons constaté que le FXII protège les neurones en culture de la mort apoptotique. Les effets bénéfiques du FXII résultent de l'interaction directe du FXII avec le récepteur du facteur de croissance épidermique (EGFR). L'activation de l'EGFR par le FXII déclenche les voies signalétiques antiapoptotiques MAPK. Il est intéressant de noter que la forme double chaîne du FXII, αFXIIa, exerce des effets protecteurs complémentaires en convertissant le précurseur du facteur de croissance hépatocytaire en sa forme mature, ce qui active à son tour le récepteur MET. Enfin, dans notre étude, nous avons observé que le FXII était exprimé dans le SNC (à la fois l'ARNm et la protéine). Cette expression est notamment retrouvée au niveau des neurones corticaux. Ce travail décrit un nouveau mécanisme d'action du FXII et décrit les neurones en tant que cellules cibles pour les effets facteur de croissance du FXII. Dans l'ensemble, ces travaux devraient donc aider à mieux comprendre comment le FXII agit dans les pathologies cérébrales en tant que sérine-protéase unique à l'interface de la thrombose, de l'inflammation et de la survie cellulaire. / Factor XII (FXII) is an 80 kDa serine protease produced and secreted by the liver that participates in the intrinsic coagulation pathway. Various studies have shown a deleterious role of FXII in cerebral pathologies like stroke, Alzheimer disease and multiple sclerosis. Nevertheless, the direct impact of FXII on neuronal cell fate remains unknown. In addition, the expression of FXII in the central nervous system (CNS) has not been investigated yet. The first objective of our work was to study the role of FXII in neuronal apoptosis and then to study the expression of FXII in the CNS. In our work, we found that FXII protects cultured neurons from apoptotic death by a growth factorlike effect. The beneficial effects of FXII result from the direct interaction with the epidermal growth factor receptor (EGFR). Activation of EGFR by FXII triggers antiapoptotic signaling MAPK pathways. Iteestigl, the to hai fo of FXII, αFXIIa, eets opleeta potetie effets oetig the hepatocyte growth factor (HGF) precursor into its mature form, which in turn activates MET receptor. Finally in our work we observed FXII expression in the CNS (both mRNA and protein). This expression is particularly found in cortical neurons. This work describes a novel mechanism of action of FXII and discloses neurons as target cells for growth factor effects of FXII. Overall, these works should thus help further understanding how FXII acts in brain diseases as a unique serine-protease at the interface of thrombosis, inflammation and cell survival.

O angioedema hereditário (AEH) é uma doença autossômica dominante, caracterizada por episódios recorrentes de edema subcutâneo, do trato gastrointestinal e das vias aéreas superiores. Em sua forma clássica, o AEH é causado por deficiência do inibidor de C1 (C1- INH) e está associado a mutações no gene SERPING1. Recentemente, foi descrito o AEH com inibidor de C1 normal, resultante de mutações no gene F12, que codifica o Fator XII da coagulação (FXII), denominado AEH-FXII; e de mutações nos genes PLG e ANGPT1, que codificam Plasminogênio e Angiopoietina-1, caracterizando os tipos AEH-PLG e AEHANGPT1. O AEH com inibidor de C1 normal não cursa com diminuição de C1-INH quantitativo e/ou funcional, ou de C4, sendo o diagnóstico realizado pela presença de sintomas clínicos sugestivos e história familiar. Os objetivos do presente estudo foram: avaliar a discriminação alélica como método alternativo ao sequenciamento por método de Sanger, para o diagnóstico de AEH com inibidor de C1 normal por mutação c.983C>A no gene F12; determinar se a discriminação alélica seria uma técnica de melhor custo-benefício; e investigar outras mutações previamente descritas no exon 9 do gene F12, e nos genes PLG e ANGPT1 em pacientes com suspeita clínica de AEH com inibidor de C1 normal, e negativos para mutação c.983C>A em F12. Cento e oitenta e quatro indivíduos incluindo 51 pacientes- índice com suspeita clínica de AEH com inibidor de C1 normal, e seus familiares, foram previamente investigados por sequenciamento de Sanger. Esses indivíduos foram investigados por discriminação alélica, e a concordância entre os resultados para a mutação c.983C>A foi verificada por estatística Kappa. Custos e tempo de execução entre os dois métodos foram avaliados. Casos-índice negativos para a mutação c.983C>A no gene F12 foram avaliados para outras mutações no exon 9 do gene F12 e para mutações previamente descritas c.988A>G e c.807G>T em PLG e ANGPT1, respectivamente, usando sequenciamento por método de Sanger. Mutação no gene F12 foi identificada em 96 indivíduos (24 pacientes- índice e 72 familiares). Em todos esses pacientes foi encontrada a mutação c.983C>A. 88 indivíduos foram negativos para esta mutação. Os resultados foram 100% concordantes entre os dois métodos. Setenta e um dos 96 pacientes positivos para mutação c.983C>A eram do sexo feminino; 78,9% e 56% eram pacientes sintomáticos do sexo feminino e masculino, respectivamente. A discriminação alélica apresentou custo e tempo de execução 79,7% e 82,3% menores em comparação ao sequenciamento por Sanger, respectivamente. Outras mutações no gene F12, e mutações em PLG e ANGPT1 não foram encontradas. Entre os pacientes negativos para mutações associadas ao AEH com inibidor de C1 normal, 11 foram diagnosticados como AEH-desconhecido e 16 como angioedema idiopático adquirido não histaminérgico. Os resultados do presente estudo nos permitiram construir um algoritmo para diagnóstico de pacientes com AEH com inibidor de C1 normal. Em áreas onde a mutação c.983C>A em F12 é predominante entre pacientes com AEH-FXII, a discriminação alélica pode ser um método adequado para screening inicial. Em pacientes com resultados negativos, sequenciamento do exon 9 de F12 pelo método de Sanger estaria indicado. Pacientes remanescentes com resultados negativos seriam genotipados para mutações previamente descritas em PLG e ANGPT1. Estudos futuros em outros locais serão necessários para estabelecer se a discriminação alélica apresentaria melhor custo-benefício, com potencial para mais acessibilidade ao diagnóstico em pacientes portadores da doença no Brasil / Hereditary angioedema (HAE) is an autosomal dominant disease characterized by recurrent episodes of edema of subcutaneous tissue, gastrointestinal tract, and upper airways. In its classical form, HAE is caused by deficiency of C1 inhibitor (C1-INH) and it is associated with mutations in the SERPING1 gene. Recently, HAE with normal C1 inhibitor has been described, resulting from mutations in F12 gene, which encodes coagulation factor XII (FXII), designated as FXII-HAE; and from mutations in PLG and ANGPT1 genes, which encode Plasminogen and Angiopoietin-1, characterizing PLG-HAE and ANGPT1-HAE types. HAE with normal C1-INH does not present with decrease in quantitative and/or functional levels of C1-INH or C4, and diagnosis is carried out by presence of suggestive clinical symptoms and family history. Diagnosis can only be confirmed by genetic sequencing. The objectives of the present study were: to evaluate the allelic discrimination as an alternative method to sequencing by Sanger method for the diagnosis of HAE with normal C1-INH due to mutation c.983C>A in F12 gene; to determine whether allelic discrimination would be a better cost-effective technique; and to investigate presence of other mutations previously described in exon 9 of F12 gene and in PLG and ANGPT1 genes in patients with clinical suspicion of HAE with normal C1-INH who were negative for c.983C>A mutation in F12. One hundred and eighty-four individuals including 51 index patients with clinical suspicion of HAE with normal C1-INH and their relatives were previously investigated by Sanger sequencing. These individuals were investigated for allelic discrimination, and concordance of the results for the c.983C>A mutation was verified by Kappa statistics. Costs and execution time of the two methods were evaluated. Negative index cases for c.983C>A mutation in F12 gene were evaluated for other mutations in exon 9 of the F12 gene and for mutations previously described c.988A>G and c.807G>T in PLG and ANGPT1, respectively, using sequencing by Sanger method. Mutation in F12 gene was identified in 96 individuals (24 index patients and 72 relatives). In all these patients, the c.983C>A mutation was found. 88 subjects were negative for this mutation. The results were 100% concordant between the two methods. Seventy-one of the 96 patients positive for the c.983C>A mutation were female; 78.9% and 56% were symptomatic female and male patients, respectively. The allelic discrimination presented cost and execution time 79.7% and 82.3% lower in comparison to sequencing by Sanger, respectively. Other mutations in F12 gene, and mutations in PLG and ANGPT1 were not found. Among patients who were negative for mutations associated to HAE with normal C1-INH, 11 were diagnosed as unknown HAE and 16 as idiopathic nonhistaminergic acquired angioedema. The results of present study allowed us to construct an algorithm for diagnosis of patients with HAE with normal C1-INH. In areas where the c.983C>A mutation in F12 is predominant among patients with FXII-HAE, allelic discrimination may be a suitable method for initial screening. In patients with negative results, sequencing of F12 exon 9 by the Sanger method would be indicated. Remaining patients with negative results would be genotyped for mutations previously described in PLG and ANGPT1. Future studies at other sites will be needed to establish whether allelic discrimination would be more cost-effective, with potential for more accessibility to diagnosis in patients with the disease in Brazil

by Chan Po Kwok. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 132-140). / Abstract --- p.i / Acknowledgement --- p.iii / Publication --- p.iv / Abbreviations --- p.v / Table of Contents --- p.vi / Chapter Chapter 1 --- Introduction / Chapter Section a --- Blood Coagulation --- p.1 / Chapter Section b --- The role of factor XII --- p.4 / Chapter Section c --- Genomic organisation of the human factor XII gene --- p.8 / Chapter Section d --- Protein structure of the human factor XII --- p.12 / Chapter Section e --- Mutations in factor XII --- p.16 / Chapter Section f --- Methods to detect mutations --- p.25 / Chapter Section g --- About the patient with factor XII deficiency --- p.29 / Chapter Section h --- Strategies used in this study --- p.30 / Chapter Chapter 2 --- Methods and Materials / Chapter Section a --- Genomic DNA extraction --- p.33 / Chapter Section b --- Polymerase chain reaction amplification --- p.34 / Chapter Section c --- Cycle sequencing --- p.35 / Chapter Section d --- Restriction enzyme digestion and cloning of PCR products --- p.36 / Chapter Section e --- Reverse transcription and polymerase chain reaction of factor XII ectopic transcript --- p.36 / Chapter Section f --- Subcloning of 95-bp novel fragment --- p.38 / Chapter Section g --- Gel mobility shift assay --- p.39 / Chapter Chapter 3 --- Results / Chapter Section a --- Analysis of the catalytic region of the human factor XII --- p.41 / Chapter Section b --- Ectopic transcript of factor XII in peripheral blood lymphocytes --- p.65 / Chapter Section c --- Analysis of 3'end untranslated region of factor XII gene --- p.70 / Chapter Section d --- Analysis of 5' flanking region of factor XII gene --- p.75 / Chapter Section e --- Analysis of intron B --- p.94 / Chapter Section f --- Analysis of 5'-b PCR product --- p.98 / Chapter Chapter 4 --- Discussions / Chapter Section a --- Mutations in the coding sequence of factor XII gene --- p.107 / Chapter Section b --- Ectopic transcript of factor XII in lymphocytes --- p.113 / Chapter Section c --- The 3'untranslated region of factor XII gene --- p.116 / Chapter Section d --- The 5'flanking region of factor XII gene --- p.117 / Chapter Section e --- Other possible explanations of undetectable factor XII mRNA --- p.120 / Chapter Section f --- The 95-bp novel sequence in ´5ة flanking region --- p.123 / Chapter Section g --- Technical difficulties --- p.126 / Chapter Section h --- Future study --- p.130 / Chapter Chapter 5 --- References --- p.132 / List of Tables --- p.137 / List of Figures --- p.138