RhD polymorphism is an evolutionary enigma since the very discovery. Theoretically, the RhD- allele carriers should be eliminated through the negative selection against RhD positive children born to RhD negative mothers. The aim of this diploma thesis was to find out whether RhD positive heterozygous genotype is associated with the reduction of latent toxoplasmosis negative effects or even associated with improved psychomotor performances, memory or higher self-confidence, intuition and cognitive performances scores. Second of all, the aim of this thesis was to find out if RhD negative homozygotes exhibit worsened performances in the abovementioned characteristics and, consequently, see whether RhD polymorphism origin and maintenance could be achieved through the heterozygote advantage. General linear models that included listed variables, RhD genotypes and phenotypes, information about toxoplasma-infection status and age as a covariate were performed. Heterozygote Advantage hypothesis was supported in operational memory models and in case of women also in psychomotor performances models. On the other hand, models of short-term memory, self-confidence, intuition and cognitive performances scores did not support the formulated hypothesis. Keywords RhD polymorphism, selection favouring heterozygotes,...

En blodtransfusion kan rädda liv men det kan också avsluta ett liv om fel blodtransfunderas. En oförenlighet mellan mottagaren och blodgivaren kan leda tillimmunisering eller hemolytiska transfusionsreaktioner (HTR) med dödlig utgång. Viktigast att ta hänsyn till är A- och B-antigenen inom AB0-systemet och D-antigenet inom Rh-systemet då dessa kan orsaka stor skada. På grund av en stor variation hos D-antigenet, i form av svagt uttryck eller skadade epitop kan individer med vissa av dessa varianter immuniseras om de erhåller RhD-positiva erytrocyter. För att minska risken för immunisering av mottagaren används därför olika reagenser för att detektera D-antigenet beroende på om individen är mottagare eller blodgivare. Syftet med studien var att utvärdera bestämning av RhD-antigen vidblodgivargruppering i en kolonn innehållande anti-D som påvisar kategori VI genom direktagglutinationsteknik (DA), samt att jämföra med befintlig metod för påvisandet avsvaga och partiella D med indirekt antiglobulintest (IAT). Tanken var att minska antaletmanuella analyser med IAT. Blodgivargruppering utfördes på helblod från 50 nyregistrerade blodgivare i AB0/D gelkort med DA-teknik. De RhD-negativa proverna analyserades med IAT för konfirmering av negativa resultat samt påvisning av RhD-varianter. Resultaten med det nya anti-D reagenset visade något starkare reaktioner än med det nuvarande reagenset. Detta kan antyda att fler skulle kunna bestämmas som RhD-positiva utan att IAT behöver utföras. Studien ger för lite underlag för några konkreta slutsatser. Avslutningsvis föreligger det ett intresse ur patientsäkerhetssynpunkt att utföra en meromfattande studie för att på djupet utvärdera bestämning av RhD-antigen meddirektagglutinationsteknik. / Blood transfusion can save lives, but if incompatible blood is transfused it could lead to immunization or fatal haemolytic transfusion reactions. The ABO and the Rh systems are considered the most clinically relevant systems. There are several variants of RhD antigen and individuals expressing some of these variants can be immunized if they receive RhD positive erythrocytes. Therefore, to reduce the risk of immunization, different reagents are used to detect the D-antigen depending on whether the individual is a recipient or a blood donor. The aim to this study was to evaluate RhD phenotyping in blood donors using a new anti-D reagent that detects partial D category VI with direct agglutination technique and to compare with the current method for detecting weak or partial D. Could the number of manual tests for the D variant with indirect antiglobulin test be reduced? Anticoagulated whole blood from 50 newly registered blood donors were grouped with the AB0/D gel card and D-negative samples was further analyzed with indirect antiglobulin test for confirmation of negative results and detection of RhD variant. Thereactions with the new anti-D showed stronger results than those from current anti-D. The results may indicate that with the new anti-D, more tests could be typed D positive, without indirect antiglobulin test. There is not enough data for proper conclusions,but there is an interest in a more comprehensive study to evaluate the D phenotyping with direct agglutination technique in the aspect of patient safety.

Une méthode de réaction de polymérisation en chaîne en temps réel (t-PCR), permettant de déterminer dès 12 semaines d'aménorrhée (SA) le statut RhD dun foetus à partir d'ADN libre dans le plasma maternel, a été mise au point dans notre laboratoire. Cette technique est appliquée en routine clinique depuis novembre 2002 aux patientes RhD négatif dans le cadre de leur suivi immuno-hématologique. Ce développement ainsi quune première évaluation portant sur 218 grossesses et 223 nouveau-nés ont été rapportés dans un travail original publié dans le J Gynecol Obstet Biol Reprod en 2005 (Minon et al. 2005). La qualité de la prédiction du statut RhD ftal a permis d'envisager l'extension du génotypage à toute patiente RhD négatif. En parallèle, différentes PCR conventionnelles ont été développées pour rechercher les gènes RHD silencieux. Lexistence de variants non fonctionnels du gène RHD - principalement trouvés dans les populations africaine et asiatique - a conduit à amplifier des segments spécifiques du gène RHD fonctionnel (séquences des exons 4 et 5) en plus dune séquence de lexon 10, afin déviter les faux positifs. Toutefois, un résultat faux positif inhabituel, lié en fait à la présence d'un greffon rénal chez l'une de nos patientes, a fait l'objet d'une publication dans Transfusion en 2006 (Minon et al. 2006). La place du génotypage foetal RHD a été réfléchie dans un contexte plus général de prise en charge et de prévention de l'alloimmunisation foeto-maternelle anti-D. Cette réflexion a permis de définir de nouvelles stratégies qui ont été décrites dans la Revue Médicale de Liège en 2006 (Minon et al. 2006). Une synthèse de notre activité tant régionale que nationale entre 2002 et 2006 a été récemment publiée dans Transfusion en 2008 (Minon et al. 2008). Elle reprend l'étude de 563 patientes et de leurs 581 nouveau-nés. Notre expérience dans les grossesses gémellaires y est rapportée. Les pièges (faux positifs et négatifs) de la technique sont abordés et des moyens de prévention pour les éviter sont proposés. Elle a été loccasion dune discussion assez exhaustive sur le génotypage RHD foetal à partir du sang maternel. La faible expression de l'antigène D chez certaines mamans pose un double problème: le premier est lié à la présence d'un gène RHD maternel "intact" qui invalide la recherche du gène RHD foetal dans le plasma. Le second est lié à la confusion induite dans lesprit de l'obstétricien par la présence d'un gène RHD maternel « normal » mais associé à une expression faible de l'antigène D, la patiente étant par ailleurs connue sérologiquement D négative. Sur ce sujet déjà épineux viennent se greffer les particularités des variants D partiels. Une nouvelle stratégie a été définie afin d'orienter l'obstétricien dans le suivi immuno-hématologique et la prophylaxie par anti-D des patientes présentant un D variant (D faible et/ou D partiel). Cette nouvelle approche concerne aussi la politique transfusionnelle à adopter en matière de sang D phéno-compatible. Cette discussion fait partie intégrante de notre sujet et reprendra notre expérience reposant maintenant sur plus de 700 déterminations de RHD ftal réalisées depuis 2007. L'extension du génotypage RHD à toutes les patientes RhD négatif encourage nos équipes à réaliser de manière systématique une prévention par immunoglobulines anti-D à 28 semaines daménorrhée (SA) chez les patientes Rh D négatif dont le foetus est RHD positif. L'introduction de cette prophylaxie modifie le suivi immuno-hématologique obstétrical traditionnel et fait l'objet d'une publication sous presse dans Acta Clinica Belgica (Minon et al. 2008). En corollaire de notre travail, nous avons démontré la fiabilité de la prédiction du sexe foetal par la recherche du gène SRY utilisé comme contrôle interne de la présence de DNA foetal dans le plasma maternel lorsque le foetus est de sexe masculin.

The boy-to-girl ratio at birth (secondary sex ratio) is around 1.06 in most populations. The sex ratio may be influenced by many factors, such as stress and immunosuppression, age of parents, parity and sex of preceding siblings. The most common human protozoan parasite in developed countries, Toxoplasma gondii (prevalence 20% - 80%), is known to change the behaviour of its intermediate hosts, thereby increasing the probability of transmission to its definitive host (the cat) by predation. The results of our retrospective cohort study suggest that the presence of Toxoplasma gondii, can influence the secondary sex ratio in humans. Depending on the antibody concentration, the probability of the birth of a boy can increase up to a value of 0.72, which means that for every 260 boys born, 100 girls are born to women with the highest concentration of anti-Toxoplasma antibodies. In accordance with results on human subjects, laboratory mice with toxoplasmosis produced a higher sex ratio than controls, in the early phase of latent infection. Our further results showed that mice in the early phase of latent infection exhibited temporarily increased production of interleukin (IL)-12 and decreased production of IL-10. The mice showed decreased production of IL-2 and nitric oxide and decreased proliferation...

Made available in DSpace on 2015-04-11T13:38:35Z (GMT). No. of bitstreams: 1 Dissertacao Francimary.pdf: 640225 bytes, checksum: 1eebacf1e67b120750607e8d1a1c46d2 (MD5) Previous issue date: 2005-10-26 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The presence of G class irregular erythrocyte alloantibodies in the blood circulation during pregnancy may cause a series of consequences to the newborn. The alloimmunization can occur under different situations, such as: blood incompatibility during delivery, miscarriage, amniocentese, blood transfusion, among others, all considered being risky. This study investigated the occurrence of regular and irregular circulating alloantibodies in women under the care of the Program of RhD-negative Expecting Mothers (PGRhN) of the Amazonas Hematology and Hemotherapy Foundation (HEMOAM). A retrospective study was carried out compiling the data of all expecting mothers under care during 2003, as well as the information regarding their newborns. A second study, prospective, was done by analyzing the blood samples of expecting mothers under the care of the PGRhN between April and November 2004, as well as the information on their respective babies. All tests were carried out following the methodology presented in the American Association of Blood Bank Technical Manual (2005). The comparison of the alloimmunization frequency between the first (5.6%) and the second (3.5%) studies showed no significant difference. The frequency of antibodies attached to the erythrocytes in the newborns of alloimmunized mothers were high, 67% in the retrospective study and 72.7% in the prospective study. The frequency of G class regular alloantibodies was 87% in the prospective study and the ABO maternal-fetal incompatibility was 17% in O type mothers with A or B type progeny. The occurrence of irregular alloimmunization in the PGRhN is still high, comparing with the numbers of other authors, indicating that the Program may be failing in preventing the problem to occur or errors may have been made during the sorotherapy of patients under risk. Also, it was observed a high frequency of G class regular alloantibodies which can cause the Hemolytic Disease of the Newborn (HDN) due to ABO incompatibility. / A presença de aloanticorpos irregulares eritrocitários da classe G, na circulação materna, pode causar serias conseqüências ao neonato. A aloimunizacao pode ocorrer em diversas situações, consideradas de risco, como partos com incompatibilidade sanguínea, aborto, amniocentese, transfusões sanguínea, dentre outras. Neste estudo foi verificada a presença de aloanticorpos circulantes regulares e irregulares, em mulheres atendidas no Programa de Gestantes Rh Negativo (PGRhN) da Fundação de Hematologia e Hemoterapia do Amazonas (HEMOAM). Foi realizado um estudo retrospectivo e para o qual, foram levantados os registros de gestantes atendidas durante o ano de 2003, como também, os dados de seus recém-nascidos. Outro estudo, prospectivo, foi realizado e foram analisadas amostras de sangue de gestantes, atendidas no PGRhN entre os meses de abril a novembro de 2004, como também de seus recém-nascidos. Todos os testes utilizados foram preconizados pelo Manual Técnico da American Association of Blood Bank (2005). A freqüência de aloimunizacao no estudo Retrospectivo (5,6%) não apresentou diferença significante com a do estudo Prospectivo (3,5%). A freqüência de anticorpos fixados as hemácias, nos recém-nascidos de mães aloimunizadas, foi bastante alta, de 67% no estudo prospectivo e de 72,7% no estudo retrospectivo. A freqüência de aloanticorpos regulares da classe G foi de 87% nas gestantes do estudo prospectivo e a incompatibilidade ABO materno-fetal foi de 17% para as mães O, com filhos A ou B. O índice de aloimunização irregular no PGRhN ainda e muito alto, quando comparado aos trabalhos de outros autores, mostrando que devem estar ocorrendo falhas na prevenção e possíveis erros na administração da soroterapia, em situações de risco. Ressalte-se ainda, a alta freqüência de aloanticorpos regulares da classe G, que podem induzir a Doença Hemolítica do recém-nascido (DHRN) por incompatibilidade ABO.

Made available in DSpace on 2014-07-11T16:47:19Z (GMT). No. of bitstreams: 2 Sá_Cynthia_iff_dou_2013.pdf: 4245727 bytes, checksum: 621887ac992f6d17a60eb59fd4d8f036 (MD5) license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) Previous issue date: 2013 / Fundação Oswaldo Cruz. Instituto Nacional de Saúde da Mulher, da Criança e do AdolescenteFernandes Figueira. Departamento de Ensino. Programa de Pós-Graduação em Saúde da Criança e da Mulher. Rio de Janeiro, RJ, Brasil. / Introdução: A Doença Hemolítica Perinatal RhD (DHPN-RhD) é decorrente da passagem transplacentária de anticorpos maternos anti Rh positivo causando hemólise no feto e recém-nascido. Este processo pode ser prevenido pela administração de imunoglobulina anti-Rh D, no entanto, quando instalada, é irreversível. Os pacientes afetados poderão desenvolver anemia e icterícia que se não tratadas adequadamente, levam a dano cerebral irreversível, clinicamente conhecido como Kernicterus ou morte. Apesar da existência de profilaxia adequada, a DHPN-RhD ainda é prevalente no Brasil, mas não é considerada para cálculo nos Estudos de Carga de Doença. Considerando que ela possa representar uma fração importante da morbidade e mortalidade perinatal e neonatal este trabalho propõe o cálculo da carga da DHPN-RhD no Brasil Objetivo: Calcular a Carga da DHPN-RhD no Brasil e Regiões do País Método: O indicador utilizado neste estudo foi o número de anos de vida perdidos ajustados por incapacidade DALY (DisabilityAdjusted Life Years). O número de DALY foi calculado a partir da soma de duas parcelas: Anos de Vida Perdidos por Morte Prematura (YLL -Years of Life Lost,) e os Anos de Vida Perdidos devido à Incapacidade (YLD - Years Lived with Disability. Os dados para o cálculo do YLL foram obtidos através do Sistema de Informação sobre Mortalidade (SIM/DATASUS). No processo de estimação do YLD, foram utilizados, como parâmetros clínicos epidemiológicos, os casos incidentes, a duração e o peso médio das incapacidades. O considerada foi de até 30 dias. Devido à gravidade do Kernicterus, considerou-se como duração 50% da expectativa de vida estimada para o País e para as macrorregiões. Para avaliar as estimativas dos pesos das incapacidades optou-se utilizar o peso 0,894 para os casos de DHPN-RhD sem sequelas e 0,459 para os casos de Kernicterus, considerando as sequelas neurológicas relacionadas ao quadro. Resultados: A DHPN-RhD possui as menores taxas de DALY em todas as regiões do País comparando com os outros eventos perinatais. As regiões Norte e Nordeste tiveram as maiores taxas para todos os eventos neonatais com exceção da DHPN-RhD cuja segunda maior taxa foi a da região Sul. Quando analisados o componente YLL, as maiores taxas estão nas regiões Norte e Nordeste e a menor no Sudeste. Em relação ao YLD, observa-se uma inversão em relação aos valores encontrados para o componente YLL, isto é, as maiores taxas foram aquelas das regiões Sudeste e Sul. Conclusão: Apesar de a Carga da DHPN-RhD no Brasil ser muito inferior aos outros eventos perinatais ele demonstra uma herança histórica de ausência de investimentos no Norte e Nordeste como demonstrado em nosso trabalho, onde o risco de morrer por DHPN-RhD é maior que nas regiões Sul e Sudeste. Assim, para o declínio da DHPN-RhD no nosso país há necessidade de melhorar a assistência perinatal e também de um reconhecimento político da contingência dessas desigualdades regionais e da necessidade de priorizar a profilaxia ao invés de tratamento. / Introduction: Rhesus Haemolytic Disease (RHD) occurs due passage of maternal antibodies anti RhD by placenta causing hemolysis in fetus and newborn. This can be prevented by administration of anti-Rh D immunoglobulin, however, when installed, is irreversible. Affected patients may develop anemia and jaundice which if not treated, lead to irreversible brain damage known as kernicterus or death. Despite the existence of appropriate prophylaxis to RhD-DHPN is still prevalent in Brazil, but is not considered for calculating the Burden of Disease Study. Whereas it may represent an important fraction of perinatal morbidity and mortality and neonatal this paper proposes calculation the burden of Rhesus Haemolytic Disease in Brazil Objective: To calculate burden of Rhesus Haemolytic Disease in Brazil and Macroregions Methods: The indicator used in this study was the number of years of life lost disability-adjusted DALY (Disability Adjusted Life Years). The number of DALYs was calculated from the sum of two components: Years of Life Lost to Premature Death (YLL-Years of Life Lost) and Years of Life Lost due to Disability (YLD - Years Lived with Disability. Data for the calculation of YLL was obtained through the Information System (SIM / DATASUL.) In the process of estimation of YLD were used as clinical epidemiological incident cases, the length and weight of disabilities. The number of incident cases calculated through analysis of the Hospital Information System of the Unified Health System (SIH-SUS). In the case of RhD without sequelae was considered the duration of up to 30 days due but to the severity of kernicterus it was considered as 50% of the duration estimated life expectancy. To evaluate the estimates of the weights of disabilities we use the weight to 0,894 cases of RHD without sequelae and 0.459 for cases of kernicterus considering neurological sequelae. Results: RHD has the lowest rates of DALYs in all regions of the country compared to other perinatal events. The North and Northeast had the highest rates for all events with the exception in the South were the rates RHD DALYS was the second highest. When analyzed the component YLL rates are the highest in the North and Northeast and the lowest rate in the Southeast. Regarding the YLD observed a reversal of the values found for the YLL component is the highest rates were those from Southeast and South Conclusion: Despite the burden of RhD Hemolytic Disease Perinatal in Brazil is much lower than the other perinatal events there is a historical legacy of lack of investment in the North and Northeast as demonstrated in our work where the risk of dying from DHPN-RhD is higher than in the South and Southeast. So for the decline of DHPN-RhD in our country there is a need to improve perinatal care and also a political recognition of the contingency of these regional inequalities and the need to prioritize the prevention rather than treatment.

This work presents the results ofour research of cell-free nucleic acids (cfNA). The first part shows changes in methylation patterns of immune response genes promoters that are detectable in plasma during the hemodialysis sessions and also differences in methylation between patients and healthy subjects. Alterations include genes that play their role in the regulation of hematopoiesis and these changes are in close relation with the need of anemia therapy. In the other plasma cfNA study we detected miRNA signatures in patients with acute myeloid leukemia at diagnosis (6 highly abundant miRNAs found) and in remission achieved after standard chemotherapy (trend to n01malization, lower levels ofthese miRNAs). Another part of work presents data from the study of potential non-invasive biomarker of bladder cancer. The amounts of cfDNA in urine are higher in patients than in healthy subjects and there were found 5 down-regulated miRNAs. Simultaneously it was established set of 30 miRNAs that are constantly present in urine supematants independently on sex, age and healthy status of subjects. The last part presents analysis ofcell-free fetal DNA. We analyzed differences between a new quantification method - droplet digital PCR and real-time PCR which is used routinely nowadays. Slightly more precise was...

Le travail présenté dans ce manuscrit traite des mécanismes d'émergence des maladies infectieuses. Une première partie du texte est consacrée à la réflexion menée sur ces processus, et à leurs conséquences pour la gestion des maladies. Dans une deuxième partie nous exposons les travaux de modélisation réalisés ; l’objectif est de tester si des changements dans l'expression d'une infection peuvent être expliqués par la dynamique de circulation de l'agent infectieux dans la population hôte, à travers la protection de groupe. Nous montrons que l'apparition de la maladie de Carré chez les lions du Serengeti peut être due à une perte d’immunité de groupe suite à l’extinction du virus. Nous étudions ensuite les conséquences sur le patron épidémique d'une protection partiellement croisée entre souches. Enfin, nous intégrons à cette problématique la structure spatiale de la population hôte, pour expliquer des observations faites sur les systèmes lagomorphes/lagovirus en France. / The study presented here aims to suggest and to test several mechanisms to explain how infectious diseases impact could increase, in relation to microparasite circulation and loss of herd protection. We first introduce the main characteristics of host/microparasite interaction and display some knowledge about emerging infectious diseases, and their consequences for diseases control. Stochastic modelling is then used to study epidemic patterns under several hypotheses and for different host/parasite systems: i) we explain canine distemper emergence in Serengeti lions through a break in virus circulation and the resulting loss of herd immunity; ii) we study the consequences of partial crossprotection induced in hosts by different strains in a small homogeneous host population; and iii) we extend this study to spatially structured host populations to explain data observed in lagomorphs/lagoviruses in France.