Nas últimas décadas, a podridão vermelha das raízes da soja (PVR), causada pelo Fusarium solani f.sp. glycines (FSG), tornou-se uma doença séria nas regiões brasileiras onde já foi constatada, sendo a utilização de cultivares resistentes um componente fundamental de um sistema integrado de controle. Este trabalho teve por objetivo pesquisar estratégias de seleção de genótipos resistentes a PVR. Várias metodologias foram implementadas: avanço de progênies até a geração F7:2 e seleção em campo infestado, de progênies selecionadas em F2. Avaliou-se o cruzamento IAC 4 x Conquista em casa de vegetação (geração F3:2) e em campo infestado (gerações F3:2, F4:2 e F5:2), através de estudo de estabilidade e adaptabilidade das progênies. As metodologias de infecção em vasos de barro e bandejas de isopor em casa de vegetação foram comparadas para melhorar a eficiência de seleção; também foi feita análise de repetibilidade dos sintomas foliares da PVR, para otimização do número de avaliações necessárias para classificar o genótipo de soja quanto à reação ao FSG. Cultivares brasileiras de soja foram avaliadas em campo infestado. Em campo, as plantas foram avaliadas no estádio R5-6, com notas variando de 1 (ausência de sintomas) a 5 (100% da raiz principal com sintomas). Em casa de vegetação, as plantas foram avaliadas aos 35 dias pós-semeadura para sintomas radiculares e da parte aérea. Houve eficiência da seleção para resistência a PVR tanto em F2 (sintomas foliares em casa de vegetação) quanto em F7:2 (sintomas radiculares em campo naturalmente infestado). A seleção praticada nas gerações intermediárias também foi eficiente para aumentar a produtividade de grãos das progênies. Para o cruzamento IAC 4 x Conquista nas gerações F3:2, F4:2 e F5:2 em campo naturalmente infestado e na F3:2 em casa de vegetação, a metodologia de Annicchiarico possibilitou a seleção de genótipos com maior estabilidade e adaptabilidade para os diferentes ambientes; com o estudo genético destas gerações, demonstrou que há efeitos gênicos aditivos e dominantes no controle genético da PVR. Além disso, constatou-se que os genes responsáveis pela resistência estão dispersos nos genitores, provavelmente agrupados em blocos gênicos. A presença de dominância indica que a seleção deve ser postergada para gerações com maior homozigose (linhagem pura); esta idéia foi reforçada pela baixa herdabilidade ao nível de plantas. O uso de bandejas de isopor foi significativamente mais eficiente do que vasos de barro para inoculação de FSG em casa de vegetação. Pela análise de repetibilidade, detectou-se que quatro avaliações foram suficientes para discriminar se o genótipo era realmente suscetível ao FSG. A correlação da geração F3:2 (IAC 4 x Conquista) entre campo infestado e casa de vegetação foi praticamente nula; no entanto, 54% das progênies selecionadas em campo também foram selecionadas em casa de vegetação. A seleção de genótipos superiores para resistência ao FSG não é tarefa fácil, mas pode ser aprimorada pelo uso conjugado de metodologias suplementares que aumentem a eficiência de seleção. / During the last decades, soybean sudden death syndrome (SDS), caused by Fusarium solani f.sp. glycines (FSG), has become a serious disease in the regions where it was encountered. The use of resistant soybean cultivars has been an important component of an integrated management system. The aim of this work was to research selection strategies of SDS-resistant genotypes. A number of methodologies were used: self pollination up to generation F7:2 and progeny selection in infested field of selected F2. The crossing IAC 4 x Conquista was studied in greenhouse (generation F3:2) and in a naturally infested field (generations F3:2, F4:2 and F5:2), where the progeny stability and adaptability study was carried out. The methodologies of infection in clay pots and polystyrene trays in greenhouse were compared to improve selection efficiency. An analysis of the repeatability of foliar SDS symptoms was also performed in order to optimize the number of evaluations needed to classify a soybean genotypes reaction to FSG. Brazilian soybean cultivars were evaluated in a naturally infested field, where plants were evaluated at stage R5-6 with scores varying from 1 (absence of symptoms) to 5 (main root 100% symptoms). In the greenhouse, the plants were evaluated 35 days after sowing for root symptoms, while the leaf symptoms was only evaluated after the appearance of symptoms. The selection done in F2 (leaf symptoms in greenhouse) as well as F7:2 (root symptoms in infested field) for FSG were efficient. The selections performed in intermediary generations were efficient in increasing the grain yield of the progeny. Studies of the F3:2, F4:2 and F5:2 generations of the IAC 4 x Conquista cross in an infested field and F3:2 in greenhouse, enabled through Annicchiaricos methodology, the selection of genotypes with greater stability and adaptability to different environments. The genetic study of these generations demonstrated there are additive and dominant genetic effects on the genetic control of SDS. Moreover, it was noted that the genes responsible for resistance are dispersed on the genitors, probably grouped into genic blocks. The presence of dominance indicates that the selection should be passed onto generations with greater homozygosity. There is also a clear indication that breeders should work with progeny lines in order to assist selection due to the low inheritability at the individual plant level. For the experiments with different FSG inoculation methodologies (clay pots and polystyrene trays) in greenhouse the use of trays was significantly more efficient; by studying the repeatability analysis, it was found that four evaluations is enough to discriminate if the genotype is really susceptible to FSG. The correlation of the F3:2 generation (IAC 4 x Conquista) between infested field and greenhouse was practically null; however, 54% of the progeny selected in the field were selected in greenhouse. As observed, the selection of superior genotypes for FSG resistance is not an easy task, but may be improved by means of new methodologies and the conjugated use of methodologies which improve selection efficiency.

A Morte Súbita dos Citros (MSC) é uma nova doença que afeta a citricultura paulista, podendo levar à morte de milhões de plantas cítricas enxertadas sobre limão 'Cravo' (Citrus limonia Osbeck), com um impacto comparável ao do vírus da Tristeza dos Citros durante as décadas de 30 e 40. Técnicas alternativas na propagação dos citros podem contornar os efeitos da MSC, seja diretamente, pela produção de subenxertos tolerantes, ou indiretamente, pelo emprego de estaquia de citros e interenxertia em combinações copa/porta-enxerto incompatíveis, mas sobre porta-enxertos tolerantes à MSC. O presente trabalho visou avaliar diferentes métodos de produção de mudas cítricas em recipientes que proporcionem possível prevenção futura das plantas à MSC em condições de campo, através de três experimentos. 1) desenvolvimento de onze porta-enxertos para fins de subenxertia, em diferentes recipientes. Os recipientes usados foram tubetes de 290 mL, sacolas de 1,7L e porta-enxertos transplantados de tubetes de 75mL para sacolas de 1,7L e 4,5L; 2) viabilização de produção de mudas de laranja 'Pêra' (Citrus sinensis L. Osbeck) sobre porta-enxertos citrumelo 'Swingle' (Poncirus trifoliata x Citrus paradisi) e limão 'Volkameriano' (Citrus volkameriana Pasquale), através de interenxertos de laranjas 'Valência' e 'Hamlin'(Citrus sinensis L. Osbeck), tangerina 'Sunki' (Citrus sunki Hort. ex Tanaka) e tangerina 'Cleópatra' (Citrus reshni Hort. ex Tanaka); 3) produção de mudas cítricas e subenxertos a partir de estacas enraizadas de citrumelo 'Swingle'. Utilizaram-se estacas herbáceas, semi-lenhosas e lenhosas obtidas de porta-enxertos de seis meses de idade, submetidas a imersão em solução com IBA 0 ou 500mg L-1 e a enxertia antes do enraizamento. Porta-enxertos produzidos diretamente em sacolas de 1,7L atingiram ponto ideal de subenxertia em menor tempo, ou seja, cerca de 100 a 150 dias após a semeadura, além de permitir a obtenção de plantas maiores e com sistema radicular adequado em mesmo espaço de tempo, quando se necessitar de subenxertos mais vigorosos no campo. A interenxertia se mostrou uma técnica promissora para propagação de citros, especialmente no caso de viabilizar o plantio de laranja 'Pêra' sobre citrumelo 'Swingle' em áreas afetadas pela MSC. Contudo, determina um ciclo de produção mais prolongado e com maior percentagem de descarte de mudas, levando-se até 17 meses para a formação da muda a partir da semeadura do porta-enxerto. Estacas herbáceas com um par de folhas são as mais indicadas para enraizamento e multiplicação de citrumelo 'Swingle'. / Citrus Sudden Death (CSD) is a new disease detected in the citrus industry in São Paulo State, Brazil. It affects all citrus varieties budded on ‘Rangpur’ Lime (Citrus limonia Osbeck), causing lost of vigor and leaf brightness, root system destruction, yield reduction, degeneration of rootstock phloem tissue and death of plants in few months. CSD has a great potential to destroy millions of plants such as Citrus Tristeza Virus has caused between 1930 and 1940. Alternative production methods of container-grown citrus nursery trees could prevent some effects of CSD, for instance, by the use of tolerant inarchings, citrus cuttings, or even interstocks in incompatible scion/rootstock combinations. This work evaluated the following alternative production methods of citrus aiming to prevent occurrence of CSD in groves: 1) production of citrus inarchings using eleven rootstocks in four container types: 290mL tubes and 1,7L and 4,5L plastic bags; 2) production of 'Pera' sweet orange (Citrus sinensis L. Osbeck) nursery trees on incompatible 'Swingle' citrumelo (Poncirus trifoliata x Citrus paradisi) and 'Volkamer' lemon (Citrus volkameriana Pasquale) with four interstocks: 'Valencia' and 'Hamlin' sweet oranges (Citrus sinensis L. Osbeck), 'Sunki' mandarin (Citrus sunki Hort. ex Tanaka) and 'Cleópatra' mandarin (Citrus reshni Hort. ex Tanaka); 3) production of citrus inarchings and nursery trees using 'Swingle' citrumelo cuttings. Herbaceous, semi-hardwood and hardwood cuttings were used and were submitted to IBA solution (0 or 500mg L-1) and budded before rooting. Production of inarchings directly sowed in 1,7L containers resulted in rootstocks ready for use about 100 to 150 days after sowing with more vigorous plants and well shaped root system. Interstocking is a promising citrus propagation technique especially regarding incompatible combinations, and could allow the use of 'Pera' sweet orange on 'Swingle' citrumelo in areas affected by CSD. On the other hand, the production cycle is longer, up to 17 months from sowing, besides producing a higher percentage of inadequate plants. Twoleaf herbaceous cuttings are indicated for appropriate rooting and multiplication of 'Swingle' citrumelo.

Embora desfrute de inegável importância econômica, os citros estão sujeitos a muitos problemas sanitários sendo alguns, limitantes para o cultivo como é o caso das doenças causadas por vírus. A morte súbita dos citros é uma doença relacionada à combinação copa/ porta-enxerto e manifesta sintomas na região da enxertia sobre porta-enxertos intolerantes. Embora sua etiologia não tenha sido determinada, há indicações que a causa da MSC esteja relacionada a uma estirpe do vírus da tristeza dos citros (CTV), a um vírus do gênero Marafivirus, ou a uma associação entre eles. Uma vez que a transformação genética têm sido considerada como uma ferramenta auxiliar a programas de melhoramento de citros, o objetivo deste trabalho foi obter plantas transgênicas de limão \'Cravo\' e laranja \'Valência\' contendo o gene da replicase do Marafivirus e estudar a regeneração e obtenção de plantas in vitro de laranja azeda, via organogênese, visando futuros trabalhos de transformação genética. Experimentos para indução da organogênese in vitro foram realizados avaliando-se citocininas (BAP, TDZ e CIN), em diferentes concentrações, isoladamente ou em combinação com ANA, condições de luminosidade (fotoperíodo de 16 h e escuro por 30 dias), meios de cultivo e explantes (provenientes de plantas germinadas in vitro e de plantas mantidas em estufa). Além disso, avaliou-se o enraizamento dos brotos regenerados. Para a transformação genética, explantes de limão \'Cravo\' e laranja \'Valência\' foram inoculados e co-cultivados com a estirpe EHA 105 de Agrobacterium tumefaciens contendo o gene da replicase do Marafivirus (em seqüência sense e antisense interligadas por um íntron). A construção gênica foi elaborada a partir do plasmídeo pCAMBIA 2201, dirigidas pelo promotor 35S e terminador NOS, contendo ainda o gene de seleção nptII. A transformação foi confirmada por análises de PCR e \'Southern blot\'. A transcrição do gene foi avaliada por RT-PCR e \'northern blot\'. A adição de BAP, combinada ou não com ANA, e em combinações com CIN ao meio de cultivo, assegurou maior formação de gemas adventícias em segmentos de epicótilo de laranja azeda. Entretanto, TDZ não se mostrou favorável a essa resposta, que também é afetada pela ausência de luz. Os explantes provenientes do cultivo in vitro mostraram-se mais favoráveis à resposta organogênica. O enraizamento das brotações de laranja azeda regeneradas foi obtido no meio MT com metade da concentração de sais, sem ou com auxinas. Foi possível obter plantas transgênicas de limão \'Cravo\' e de laranja \'Valência\' contendo o gene da replicase do Marafivirus utilizando-se segmentos internodais como explantes. A análise de \'Southern blot\' confirmou a integração de um a quatro eventos de inserção do transgene no genoma das plantas. A transcrição do gene da replicase do Marafivirus e do gene nptII foi observada por RT-PCR. / In spite of great economic importance, the citrus industry is affected by many phytopathological problems some, limiting its cultivation such as virus-caused diseases. The citrus sudden death disease is related to scion/rootstock combinations and manifests symptoms in the grafting area of intolerant rootstocks. Although its etiology has not been determined, there are indications that the cause of MSC might be related to a strain of the Citrus tristeza virus (CTV), to a virus of the Marafivirus group, or to an association of both viruses. Since the genetic transformation has been considered as an auxiliary tool to programs of citrus improvement, the objectives of this work were to obtain transgenic plants of the Rangpur lime and Valencia sweet orange containing the Marafivirus replicase gene and study the in vitro regeneration of sour orange plants through organogenesis, aiming for future work in genetic transformation. Experiments for induction of in vitro organogenesis were carried out evaluating citocinins (BAP, TDZ and KIN), in different concentrations, separately or in combination with NAA, lighting conditions (photoperiod of 16 hours and darkness for 30 days), cultivation media and explants (coming from in vitro germinated plants and from green house cultivated plants). Besides this, rooting of the regenerated shoots was evaluated. For the genetic transformation, Rangpur lime and Valencia sweet orange explants were inoculated and co-cultivated with the EHA-105 Agrobacterium tumefaciens strain containing the Marafivirus replicase gene (in sense and antisense sequence linked by an intron). The genetic construct used derived from the pCAMBIA 2201 plasmid, driven by the 35S promoter and NOS terminator, containing the selection nptII gene. The genetic transformation was confirmed by PCR and Southern blot analysis. The gene transcription was evaluated by RT-PCR and northern blot. The addition of BAP to the culture medium, combined or not with NAA, and in combinations with KIN, assured a greater formation of adventitious buds in sour orange epicotyl segments. However, TDZ was not favorable to this response, that is also affected by the absence of light. Explants coming from in vitro cultivation were more favorable to the organogenic response. Rooting of sour orange regenerated shoots was obtained in MT medium with half the salt concentration, with or without auxin. It was possible to obtain transgenic Rangpur lime and Valencia sweet orange plants containing the Marafivirus replicase gene using internodal segments as explants. The Southern blot analysis confirmed the integration of one to four copies of the transgene in the plant genome. The transcription of the Marafivirus replicase gene and the nptII gene was observed by RT-PCR.

Work name: The analysis of the Health Rescue Service City of Prag trips for the period 2007 - 2009 Aim of work: The aim of this work is to describe and assess by graphs the analysis of the Health Rescue Service City of Prag trips in Prag for the period 2007 - 2009. Further to classify its historical progress, describe its current state including the modern equipment and also describe access of resuscitations for the same period. Method: In this work a method of data collection from avaiable literature of the Health Rescue Service City of Prag is used as well as the consultation with experienced rescue workers and people who work on the operation centre and who are processing the data and statistics of the Health Rescue Service City of Prag trips. Results: The result of this work is the graphical representation of the single trips for the urgent life - threatening events and their subsequent comparison for the period 2007 - 2009. There is also the evaluation of the success of interventions. Key words: the Health Rescue Service, the medical emergency, a rescue worker, an emergency event, a sudden cardiac arrest, the resuscitation

A Morte Súbita dos Cítros (MSC) é uma doença que afeta plantas de laranjeira-doce enxertadas em limão Cravo ou Volkameriano e ainda não tem o agente causal determinado. Para entender quais são as principais mudanças fisiológicas causadas pela MSC, analisou-se as alterações nos metabolitos primários, tanto da casca da copa quanto do cavalo. Fez-se análises dos extratos hexânico e alcoólico com RMN em alta resolução e do extrato hexânico com cromatografia gasosa. Também analisou-se os metabolitos diretamente nas cascas com técnicas de RMN em alta e baixa resolução. Com essas análises foi possível observar que a MSC altera a deposição de triacilglicerídeos e sacarose nas cascas. A MSC também modifica o perfil de ácidos graxos, com o decréscimo dos teores de ácido oléico e linolênico e aumento dos ácidos cáprico, láurico, mirístico, plamítico, esteárico e linoléico, com a evolução da doença. Com esses resultados foi possível entender as principais alterações bioquímicas causadas pela MSC, como também demonstrar que elas podem ser usadas de forma complementar no diagnóstico da doença. Dentre todos os métodos avaliados, as análises das cascas com RMN em baixa resolução, com as técnicas CPMG e de precessão livre em onda contínua (CWFP), foram as mais rápidas (em alguns segundos) e eficientes para discriminar as plantas assintomáticas das sintomáticas. A discriminação das plantas pelos dados de RMN foi realizada com métodos quimiométricos como análise de componentes principais (PCA), análises de agrupamentos hierárquicos (HCA) e regressão por mínimos quadrados parciais (PLS). / Citrus Sudden Death (CSD) is a new graft-transmissible disease of sweet orange grafted on Rangpur lime and Citrus volkameriana rootstocks. The causal agent is unknown. To understand the main physiological changes caused by the CSD, we analyzed the variations in the primary metabolites content in the bark of scion and rootstock. The hexanic and hydroalcoholic extracts were analyzed by high resolution NMR. The hexanic extracts were also analyzed by gas chromatography. The metabolites were also analyzed directly in the barks with high and low resolution NMR techniques. With these analyses it was possible to observe that CSD modifies the content of triglycerides and sucrose in the barks. The disease also changes the fatty acids profile, with a decrease in the oleic and linolenic and an increase in the capric, lauric, miristic, palmitic, stearic and linoleic content. With these results it was possible to understand the main biochemical disorders caused for the CSD, as well as to demonstrate that they can be used as complementary information in the disease diagnosis. The analysis of the barks with low resolution NMR techniques CPMG and continuous wave free precession (CWFP), had been the fastest (few seconds) and the most efficient one to discriminate between the symptomless and symptomatic plants. The discrimination of the plants by NMR data had been carried with chemometric methods such as principal component analysis (PCA), hierarchic cluster analysis (HCA) and partial least square regression (PLS).

La mort subite du nourrisson (MSN) est un phénomène imprévisible et mal compris. Elle est définie par « le décès d’un nourrisson âgé de moins d’un an dont les causes restent inexpliquées malgré des investigations approfondies, incluant une autopsie et un examen de la scène du décès ». Si l’incidence est en décroissance depuis 1994 et le début des campagnes de prévention, la MSN reste la première cause de décès post-néonatale. La MSN est un phénomène multifactoriel qui survient préférentiellement chez des enfants vulnérables exposés à des facteurs de risques environnementaux. Ainsi, le fait d’identifier ces enfants à risque est un enjeu majeur dans la prévention de cette pathologie. La réponse vagale exacerbée, exprimée biologiquement par une surexpression des récepteurs muscariniques de sous-types M2 (RM2), pourrait être un facteur de risque de MSN. Nous avons déjà constaté cette anomalie biologique dans notre modèle animal d’hyperréactivité vagale et dans des cœurs de nourrissons décédés de MSN. Le but de ces travaux est d’analyser l’expression sanguine des RM2 en tant que paramètre biologique reflétant une hyperactivité vagale chez l’Homme, dans des cas de malaises vagaux et de malaises sévères inexpliqués du nourrisson. L’implication de la génétique a également été étudiée dans une famille présentant plusieurs cas de MSN. Une surexpression des RM2 a été observée chez des patients présentant des malaises vagaux. Pour la première fois, des anomalies biologiques ont été identifiées dans cette pathologie. Si l’on parvenait à valider ce paramètre biologique en tant que marqueur de risque, cela pourrait permettre d’aider au diagnostic différentiel et à la prise en charge thérapeutique de ces syncopes vagales. Les mêmes anomalies ont été observées chez des enfants de moins d’un an présentant des malaises sévères idiopathiques. Après une première analyse des données du séquençage haut débit d’exomes issus d’une famille présentant plusieurs cas de MSN, nous avons pu identifier 3 gènes pouvant être impliqués dans la MSN. Néanmoins, le scénario et le mode de transmission sont difficiles à définir. Les premières hypothèses s’orientent vers un digénisme ou même un trigénisme. La surexpression des RM2 chez les 2 parents et chez certains enfants est un premier élément suggérant la transmission du caractère « hyperactivité vagale » chez l’Homme.Nos travaux ont permis de mettre en évidence une anomalie biologique commune entre les malaises vagaux, les malaises inexpliqués du jeune enfant et la MSN, à savoir la surexpression des RM2. Ce paramètre, facilement dosable dans le sang, pourrait être un élément complémentaire dans le diagnostic différentiel et la prise en charge de ces pathologies, notamment chez les jeunes enfants pour lesquels ces malaises peuvent être très délétères. L’avancée dans le séquençage du génome permettra peut-être l’identification de facteurs de risque génétiques impliqués dans les malaises inexpliqués ou les MSN. / Sudden Infant Death Syndrome (SIDS) is an unpredictable and poorly understood phenomenon. It is defined as the "sudden unexpected death of a child younger than one year during sleep that cannot be explained after a postmortem evaluation including autopsy, a thorough history, and scene evaluation". Although the incidence has been decreasing since 1994 and the start of prevention campaigns, SIDS remains the leading cause of post-neonatal death. SIDS is a multifactorial phenomenon that occurs preferentially in vulnerable infants exposed to environmental risk factors. Thus, identifying these children at risk is a major challenge in the prevention of this pathology. The exacerbated vagal response, biologically expressed by overexpression of muscarinic M2 receptors (M2R), may be a risk factor for SIDS. We have already observed this biological abnormality in our animal model of vagal hyperreactivity and in hearts of SIDS. The aim of this work is to analyze the blood expression of M2R as a biological parameter reflecting vagal hyperreactivity in humans, in cases of reflex syncope and idiopathic apparent life-threatening events (iALTE) of infants. The involvement of genetics has also been studied in a family with several cases of SIDS (SIDS family). Overexpression of M2R has been observed in patients with reflex syncope. For the first time, biological abnormalities have been identified in this pathology. If this biological parameter could be validated as a risk marker, it could help for differentially diagnosis and treatment of these vagal syncopes. The same abnormalities were observed in children under one year old with iALTE. After a first analysis of the data of the “next generation sequencing” of the exomes of our “SIDS family”, we were able to identify 3 genes that could be involved in SIDS. However, the scenario and the mode of transmission are difficult to define. The first hypotheses are oriented towards a digenism or even a trigenism. The overexpression of M2R in both parents is a first element suggesting the genetic transmission of the character "vagal hyperactivity" in humans. Our work highlights a biological abnormality which is common to reflex syncope, iALTE and SIDS, namely the overexpression of M2R. These results confirm the hypothesis of the involvement of the vagal system overactivity in these pathologies. This parameter, easily measurable in the blood, could be a complementary assessment useful in the differential diagnosis and the management of these pathologies, in particular in infants for whom syncope can be very harmful. The development of the sequencing of human genome will probably allow the identification of genetic risk factors involved in iALTE or SIDS.

Sudden cardiac death (SCD) in a young person is a rare but tragic event, and the potential of prevention is unknown. The aim of this thesis is to contribute to the prevention by analysing SCD in the young in Sweden during the period 1992-1999. Data of SCD in the young based on a national registry is not previously reported. The approach is broad, covering the spectrum from epidemiology to supportive needs of families confronted with SCD. The survey methods comprised analyses of national registries, questionnaires, personal interviews, forensic-, police-, medical- and military conscription records. The SCD group selected from the database of the National Board of forensic Medicine consisted of 181 persons, 15 to 35 years old, who had suffered an SCD during 1992-1999 in Sweden, 132 men (73 %) and 49 women (27 %). The mean incidence was 0.93 per 100,000 per year. The trend showed no decrease during the surveyed years, 1992-1999. The most common diagnoses were the structurally normal heart (21 %), coronary artery disease (18 %), and dilated cardiomyopathy (12 %). In a study group of 162 individuals (19 cases of aortic aneurysm, 17 men and two women, were excluded), ECGs, symptoms and lifestyle factors were analysed and related to the autopsy findings. ECGs were available in 66 individuals (59 men and seven women) and 50 % of these were pathological. The most frequent aberrations were repolarisation abnormalities and in half of the cases with more than one ECG a development in a pathological direction was seen. In four out of ten seeking medical advice because of symptoms an ECG was taken and three of these were pathological. Possibly cardiac-related alpitations were common, but also non-specific symptoms such as fatigue after an influenza- like illness. It was not possible to link a certain sign or symptom to a specific diagnosis. In 26 (16 %) there was a family history of SCD. Physical activity and body mass index (BMI) in men were the same as in a control group, whilst women had a higher BMI and a lower level of physical activity than the controls. In coronary artery disease deaths there were a high percentage of smokers and BMI was higher than in the controls in both sexes. Competing athletes more often died during physical activity than non-athletes, but were not overrepresented in the SCD group. The majority of the athletes who died during physical activity had an underlying structural cardiac disease. Death during sleep was the most common mode of death in subjects with structurally normal heart. A lack of supportive structures in the handling of bereaved relatives were disclosed in the interviews. Most participants felt that they had been left mainly to themselves to find information and support. A common reflection from the bereaved was that there is a need of the same supportive routines in cases of a single death as in accidents where there are several casualties. The bereaved had a need of getting an explanation and a need of supportive structures. The cognitive dimension of understanding and the emotional dimension of being understood were found to be significant for the complex processes of mourning and recreating one’s life as a bereaved. In summary, SCD was uncommon in the young, but the incidence was not decreasing during the study period. The most common autopsy findings were the structurally normal heart and coronary artery disease. Symptoms preceding SCD were common but often misinterpreted. The SCD group was very similar to the normal population with regard to life style factors. In certain cardiac disorders physical activity seemed to trigger sudden death, whilst in others death during sleep was the most common mode of death. There is no single test which predicts if a person is at risk of SCD. A further cardiac evaluation in cases with pathological ECGs, and in cases with a positive family history or serious unexplained symptoms such as syncope, might permit the early identification of persons at risk of SCD. ECG is an underused tool in the investigation of symptoms, and a database with old ECGs available for comparison could be useful in the prevention of SCD. There is a need of better care of the bereaved, and based on our findings we propose the introduction of a supportive program.

Der Hochwasserschutz stellt für die gegenwärtige Gesellschaft eine Thematik von zunehmender Aktualität dar, besonders unter Berücksichtigung der Verschärfung der extremen meteorologischen Ereignisse infolge des Klimawandels einerseits und der Vergrößerung des Schadenpotenzials in den Auengebieten andererseits. Zu den möglichen Schutzmaßnahmen im Flussmittellauf zählt die in ökologischer Hinsicht sehr effektive Deichrückverlegung (DRV). Um ihre Wirksamkeit bei der Dämpfung der Hochwasserspitzen zu untersuchen, wird eine Parameterstudie mit einem eindimensionalen hydronumerischen Modell (MIKE11) durchgeführt. Dabei wird besonderes Augenmerk auf die Hüllkurve der sich einstellenden Wassertiefe gerichtet, sowohl oberhalb als auch innerhalb bzw. unterhalb des DRV-Abschnitts. Zunächst wird der Effekt einer Flussaufweitung bei stationärer Strömung theoretisch analysiert. Ausgehend von den Strömungseigenschaften und vom Breitenverhältnis wird eine Bestimmungsgleichung für die effektive Mindestlänge LAeff einer Aufweitung abgeleitet, ab der eine Wasserspiegelabsenkung stromauf hervorgerufen wird. Die darauffolgenden Untersuchungen beschreiben die Auswirkungen einer solchen Maßnahme bei instationärem Abfluss auf zwei ausgewählte synthetische Hochwasserwellen unter jeweils vier verschiedenen Ansätzen zur Erfassung einer Aufweitung. Die Beiträge zur Retention vom Hauptgerinne und von der Querschnittsvergrößerung werden getrennt berücksichtigt. Darüber hinaus werden Überlegungen über das Verhalten von Hochwasserwellen mit gleichem Abflussscheitel aber unterschiedlicher Fülle beim Durchlaufen der Flussaufweitung angestellt. Der Gültigkeitsbereich des stationären Ansatzes für die eindimensionale numerische Simulation einer DRV wird mithilfe eines Kriteriums definiert, das auf einer für die Abminderung des Wassertiefenscheitels im Hauptgerinne semi-empirisch abgeleiteten Formel und auf einer Abschätzung der Retention in der DRV basiert. Die Anwendung des o. g. Kriteriums auf die betrachteten Hochwasserereignisse zeigt, dass eine stationäre Betrachtungsweise lediglich für extrem flache Hochwasserwellen (ab einer Anstiegszeit von einigen Tagen) gerechtfertigt ist. Durch die Ergebnisauswertung von 960 Kombinationen der geometrischen bzw. hydrodynamischen Parameter bei einer DRV im Hochwasserfall konnte der Zusammenhang zwischen den einzelnen Kenngrößen (Länge und Breite der DRV, Rauheit im Hauptgerinne bzw. im Vorland, Hochwasserganglinie) und der Auswirkung der DRV auf Wasserstand und Durchfluss hergestellt werden. Während oberhalb einer DRV mit einer Länge größer LAeff immer mit einer Wasserspiegelabsenkung zu rechnen ist, profitieren die Unterlieger von dieser Schutzmaßnahme nur bei kleineren Hochwasserereignissen oder bei Deichrückverlegungen mit erheblicher Ausdehnung. Dank einer gezielten Auswahl der Ausgangsgeometrie lassen sich die numerischen Ergebnisse auch auf den Elbabschnitt bei Dresden als Anhaltswerte übertragen. / Flood protection is a topic of increasing urgency in today’s society considering two main factors. Firstly, the growth in damage potential on the riparian zones and secondly, the increase in extreme meteorological events due to climate change. Among the possible flood protection measures for a river’s middle reach, dike relocation (DRV) is especially effective from an ecological point of view. To investigate its effectiveness in flood peak attenuation, a parametric study is performed using a one-dimensional hydro-numerical model (MIKE11). Special attention is given to the envelope of the resulting water depths upstream, downstream and within the DRV stretch. Initially, the effects of river enlargement under steady flow conditions are analysed theoretically. Key hydraulic parameters and the width ratio are used in deriving an equation to determine the minimum effective length LAeff of an enlargement that will reduce the upstream water depth. An investigation of an enlargement in unsteady flow conditions is then made, focussing on two specific synthetic flood waves with four different models for the enlargement. The contributions to discharge retention from the main channel and the cross section expansion are successfully separately examinated. Considerations are then made for the behaviour of flood waves passing the river enlargement with the same peak discharge but different volumes. A criterion for the validity range of the steady flow approach of the one-dimensional, numerical simulation of a DRV is then defined. This is based on a semi-empirically derived formula for water level attenuation in the main channel, and an estimate of discharge retention in the DRV. The application of this criterion to the observed flood events shows that the steady flow assumption is only valid for extremely flat flood waves (with time to peak of a few days or more). An analysis of the results from 960 combinations of geometric and hydraulic parameters for the DRV during a flood event is then made. This allowed to derive the relation between individual characteristics (length and width of the DRV, roughness in the main channel and flood plain, flood hydrograph) and the effect of the DRV on water level and discharge. Whilst one can always expect a decrease in the water level upstream of a DRV with length greater than LAeff , the riverside dwellings downstream would only benefit in small scale flood events or with a dike relocation of extensive dimensions. Thanks to the purposefully selected initial geometry, the numerical results can now provide reference values for Dresden’s reach of the river Elbe. / La difesa dalle piene rappresenta per la società odierna una tematica di crescente attualità se si considera in particolare da una parte la crescita del potenziale di danno nelle zone di pertinenza fluviale e dall’altra l’acuirsi di eventi meteorologici estremi dovuto al cambiamento climatico in atto. Tra i possibili interventi di protezione dalle piene nei fiumi di pianura si colloca l’espansione golenale, misura che risulta oltretutto assai efficace dal punto di vista ecologico. Al fine di indagarne anche l’efficacia nei confronti della laminazione del colmo di piena si effettua uno studio parametrico con un programma di modellazione idrodinamica monodimensionale (MIKE11), ponendo particolare attenzione all’inviluppo dei tiranti nel tratto dell’espansione, nonché nei tratti confinanti rispettivamente a monte e a valle della stessa. Dapprima si analizza dal punto di vista teorico l’effetto di un allargamento fluviale in moto stazionario. A partire dalle condizioni idrauliche della corrente e dal rapporto di restringimento si ricava una formula per la lunghezza minima efficace LAeff dell’allargamento, a partire dalla quale si genera un abbassamento del pelo libero nel tratto di monte. Nella successiva analisi di un allargamento in moto vario se ne studia l’effetto su due onde di piena sintetiche e per quattro differenti schematizzazioni dell’allargamento, considerando separatamente i contributi alla laminazione dovuti all’alveo principale e all’allargamento della sezione. Ciò consente tra l’altro di trarre conclusioni sul comportamento di onde di piena aventi la stessa portata al colmo ma differente volume al passare per l’allargamento. Per mezzo di un criterio basato su una formula semiempirica della riduzione del tirante al colmo per l’alveo principale e su una stima della laminazione all’interno dell’espansione golenale, sviluppate nell’ambito di questo lavoro, é possibile definire il campo di validità dell’approccio stazionario alla simulazione monodimensionale di un’espansione golenale. L’applicazione di tale criterio agli eventi di piena presi in considerazione rivela che l’approccio stazionario si giustifica solo per onde di piena estremamente piatte (tempo di crescita a partire da alcuni giorni). Attraverso l’analisi dei risultati di 960 combinazioni dei parametri geometrico-idraulici di un’espansione golenale al passaggio di un’onda di piena si è potuto ricavare il rapporto tra le singole grandezze significative (lunghezza e larghezza dell’espansione golenale, scabrezza dell’alveo e della golena, idrogramma dell’onda di piena) e l’effetto dell’espansione sui tiranti e sulle portate. Se a monte di un’espansione golenale di lunghezza superiore a LAeff si presenta sempre una abbassamento del pelo libero, a valle della stessa si può trarre profitto da un simile intervento solo nel caso di onde di piena modeste o per espansioni golenali di dimensioni notevoli. Una scelta mirata della geometria iniziale consente di estendere i risultati numerici, quali valori indicativi, al tratto del fiume Elba all’altezza di Dresda.

Background - Regular exercise reduces the risk of cardiovascular disease and subsequent sudden cardiac death (SCD). However, a small, but notable proportion of athletes die suddenly due to inherited or congenital disorders of the heart that predispose to malignant ventricular arrhythmias. Such tragedies are highly publicised, particularly when high-profile athletes are involved. To date, limited evidence for the efficacy of cardiovascular pre-participation screening exists outside of the Italian experience. Furthermore, limited data exists examining the impact of ethnicity on cardiac adaptations to physical training. Whilst the cardiovascular benefits of exercise are well known, the impact of life-long endurance exercise is less well understood. Long term high-intensity endurance exercise is associated with changes in cardiac morphology together with electrocardiographic alterations that are believed to be physiologic in nature. Recent data however, has suggested a number of deleterious adaptive changes in cardiac structure, function and electrical activity in response to life-long endurance activity. Aims and Objectives - The aims of this PhD were; 1) To find an effective preparticipation screening method that would successfully identify pre-existing cardiovascular abnormalities, 2) To identify the prevalence of hypertrophic cardiomyopathy and Long QT syndrome in elite UK athletes; 3) To examine the impact and significance of ethnicity upon left ventricular remodelling in elite athletes, and 4) To examine the acute and chronic impact of ultra-endurance exercise across the life-span in male endurance athletes. Major Results and Conclusions – 1) Study 2 sought to confirm the efficacy of resting 12-Lead ECG ‘alongside’ personal/family history questionnaires and physical examinations as collective tools to identify diseases that have the potential of causing sudden death within a cohort of elite junior athletes (n=1074) and physically active school children (n=1646). Nine participants were identified with a positive diagnosis of a disease associated with SCD. None of those diagnosed with a disease associated with SCD were symptomatic or had a family history of note. Thus, personal symptoms and family history questionnaires alone are inadequate in the identification of individuals with diseases associated with SCD. In conclusion, resting 12-Lead ECG is paramount when screening for diseases that have the potential of causing sudden death in the young. 2) Study 3 examined 3,500 asymptomatic elite athletes (75% male) with a mean age of 20.5 ± 5.8 years with 12-lead ECG and 2-dimensional echocardiography. None had a known family history of HCM. Of the 3,500 athletes, 53 (1.5%) had LVH (mean 13.6 ± 0.9mm, range 13 to 16mm), and of these 50 had a dilated LV cavity with normal diastolic function to indicate physiological left ventricular hypertrophy. Three (0.08%) athletes with LVH had a non-dilated LV cavity and associated deep T-wave inversion that could have been consistent with HCM. However, none of the 3 athletes had any other phenotypic features of HCM on further non-invasive testing and none had first-degree relatives with features of HCM. In conclusion, the prevalence of HCM in elite athletes is significantly less than in the general population; with the demands of strenuous exercise on the cardiovascular system selecting out most individuals with HCM. Study 4 examined 2000 elite athletes in order to identify the prevalence of Long QT syndrome. Three athletes had a QTc value of >500 ms and all exhibited one of: paradoxical prolongation of QTc during exercise, a confirmatory genetic mutation, or prolonged QTc in a first-degree relative. In contrast, none of the athletes with a QTc value of <500 ms had any other features to indicate LQTS. Accordingly, the prevalence of a prolonged QTc interval in elite British athletes is 0.4%. 3) Study 6 examined 300 nationally ranked UK black male athletes (mean age 20.5 years) in comparison to 150 black and white sedentary individuals and 300 highly-trained white male athletes. Black athletes exhibited greater LV wall thickness and cavity size compared with sedentary black and white individuals. Black athletes had greater LV wall thickness compared with white athletes. A minority of black athlete’s exhibit LVH ≥15 mm; proposing that in the absence of cardiac symptoms or a family history of HCM, an LV wall thickness ≥15 mm in black athletes may represent physiologic LVH when the LV cavity is enlarged and diastolic indexes are normal. 7 black athletes (12%) with LVH displaying deep T-wave inversions in leads V1 to V4. In conclusion, in the absence of obvious pathology, these electrical anomalies in black athletes likely represent a normal spectrum of ECG changes in response to physical training. 4) Study 8 examined 17 male participants (age 33.5 ± 6.5 years, 26–40 years) using cardiac magnetic resonance (CMR) and echocardiography before and after a marathon to investigate the relationship between systolic function and diastolic function against biomarkers of cardiac damage. Results demonstrates biomarkers of myocardial cell damage following an acute bout of prolonged exercise are not associated with either systolic or diastolic functional measures, and do not seem to be associated with any detectable myocardial inflammation, oedema, or scarring using either gold standard techniques of gadolinium enhanced CMR or echocardiography respectively. The impact of multiple episodes of prolonged exercise, as experienced by highly trained veteran endurance athlete is not fully understood. 5) Study 10 examined the cardiac structure and function of 12 life-long, competitive endurance veteran athletes (> 50 yrs, mean ± SD marathons 178 ± 209 (range 20 – 650)) against 17 young male endurance athletes (<40 yrs) using echocardiography and CMR with late gadolinium enhancement (LGE) to assess myocardial fibrosis. Lifelong veteran athletes had smaller LV and RV end-diastolic and end-systolic volumes (p<0.05) but maintained LV and RV systolic function compared with young athletes. In 6 (50%) of the veteran athletes LGE of CMR indicated the presence of myocardial fibrosis; no LGE in the young athletes. The prevalence of LGE in veteran athletes was not associated with the number of competitive marathons or ultra-endurance marathons (>50 miles) completed, age, LV and RV end-diastolic volumes or LV mass (p>0.05). In conclusion, there is limited evidence at present demonstrating that cardiovascular re-modelling following lifelong endurance exercise leads to long-term disease progression, cardiovascular disability or SCD.

It is well known that most fluid flows observed in nature or encountered in engineering applications are turbulent and involve separation. Fluid flows in turbines, diffusers and channels with sudden expansions are among the widely observed areas where separation substantially alters the flow field and gives rise to complex flow dynamics. Such types of flows are referred to as internal flows since they are confined within solid surfaces and predominantly involve the generation or utilization of mechanical power. However, there is also a vast variety of engineering applications where the fluid flows past solid structures, such as the flow of air around an airplane or that of water around a submarine. These are called external flows and as in the former case the downstream evolution of the flow field is crucially influenced by separation. The present doctoral thesis addresses both internal and external separated flows by means of direct numerical simulations of the incompressible Navier-Stokes equations. For internal flows, the wall-driven flow in a onesided expansion channel and the pressure-driven flow in a plane channel with a single thin-plate obstruction have been studied in the fully developed turbulent state. Since such geometrical configurations involve spatially developing turbulent flows, proper inflow conditions are to be employed in order to provide a realistic fully turbulent flow at the input. For this purpose, a newly developed technique has been used in order to mimic an infinitely long channel section upstream of the expansion and the obstruction, respectively. With this approach, we are able to gather accurate mean flow and turbulence statistics throughout each flow domain and to explore in detail the instantaneous flow topology in the separated shear layers, recirculation regions as well as the recovery zones. For external flows, on the other hand, the flow past a prolate spheroid has been studied. Here, a wide range of Reynolds numbers is taken into consideration. Based on the characteristics of the vortical structures in the wake, the flow past a prolate spheroid is classified as laminar (steady or unsteady), transitional or turbulent. In each flow regime, the characteristic features of the flow are investigated by means of detailed frequency analysis, instantaneous vortex topology and three-dimensional flow visualizations.