Este estudo teve como objetivo compreender o significado de ser pai de criança com estomia. Para tal compreensão, foram utilizados o referencial teórico da antropologia interpretativa de Geertz e o método etnográfico. Participaram deste estudo sete pais de criança com estomia, cujos filhos são cadastrados nos Serviços de Assistência às Pessoas com Estomia no Sul de Minas Gerais. As questões éticas foram respeitadas e os informantes assinaram o Termo de Consentimento Livre e Esclarecido para participarem do estudo. A coleta de dados ocorreu no período de abril a agosto de 2013, no domicílio dos pais por meio de entrevistas semiestruturadas gravadas, de observação participante e de anotações em um diário de campo. Da análise, surgiu um eixo central transversal que é "a informação e desinformação" que é transpassado por outro eixo perpendicular que o divide em dois momentos: "meu filho está adoecendo: o que fazer" e "a experiência de ter um filho com estoma: como conviver". O primeiro momento revela que os pais, em busca do diagnóstico de seu filho, depararam-se com a precariedade dos serviços de saúde, com a desinformação e o despreparo dos profissionais de saúde, com filas imensas para a realização de exames e a com dificuldade para consolidar o diagnóstico, o que representou uma verdadeira peregrinação pelo sistema de saúde. Depois de confirmado o diagnóstico e a cirurgia, os percalços se instalaram pouco a pouco e, junto a eles, o aparecimento de sentimentos como medo, tristeza e ansiedade pela nova condição em sua família. No segundo momento são abordados a alta hospitalar, o retorno ao lar com a criança com estomia, o convívio social, as atividades escolares e as dificuldades e os conflitos enfrentados. A doença apresenta-se para a família como reformuladora de papeis. Assim, por meio deste estudo identificamos que a nova condição que se instala no lar, faz com queo pai comece a repensar sobre o seu papel e se inicia uma divisão de tarefas com a mãe em relação à higiene, à troca de equipamento coletor, à alimentação e à vida escolar da criança. É de suma importância a inserção de disciplinas como a antropologia e a sociologia da saúde na dinâmica curricular dos cursos de graduação na área de saúde, para sensibilizar os futuros profissionais no sentido de melhor compreensão do homem como um ser singular que carrega consigo experiências, crenças e cultura. No entanto, em muitas universidades estas disciplinas são ministradas no início dos cursos de graduação e acabam tornando-se distantes das práticas acadêmicas, nas quais o modelo biomédico é privilegiado. É premente a necessidade de conjugar saberes para tratar o ser humano, pois a dimensão da doença não se restringe à esfera biológica. O processo de comunicação deve ser repensado, pois pude apreender que a informação é dada pelo profissional de saúde, todavia os pais não a compreendem. Entendo que a Enfermagem deve repensar o planejamento da assistência à criança com estomia, incluindo o pai nesse contexto. Reitero a necessidade do envolvimento da enfermagem no acompanhamento da criança e dos pais e a readequação da escola tornando-a mais inclusiva / This study aimed to understand the meaning of being a parent of a child with ostomy. To achieve this understanding, we used the theoretical framework of interpretive anthropology of Geertz and the ethnographic method. Participated in this study seven parents of child with Ostomy, whose children are registered in the services of assistance to People with Ostomy in southern Minas Gerais. Ethical issues were respected and informants signed an informed consent to participate in the study. Data collection occurred in the period from April to August of 2013, at the domicile of the parents by semi-structured interviews, participant observation and recorded notes in a field notebook. From the analysis, emerged a transverse axis that is "the information and misinformation" which is wounded by another perpendicular axis that divides into two moments: "my son is sick: what to do" and "the experience of having a child with Stoma: how to live". The first time reveals that parents in search of the diagnosis of their sons encounter the precariousness of health services, with misinformation and preparation of health professionals, with huge queues for performing tests and with difficulty to consolidate the diagnosis, which represented a real pilgrimage by the health system. After confirmed the diagnosis and surgery, the mishaps settled gradually and, together with them, the emergence of feelings such as fear, sadness and anxiety for the new condition in their family. In the second time are approached the hospital discharge, the return home with the child with Ostomy, social conviviality, school activities and the difficulties and conflicts faced. The disease presents itself to the family as reformer of roles. Thus, by this study we identified that the new condition that is installed in the home, makes the father begins to rethink his and then starts the division of labour with the mother in relation to hygiene, exchange of collector equipment, food and to the school life of the child. It is very important entering disciplines such as anthropology and sociology of health in the curriculum of undergraduate dynamics in healthcare, to sensitize the future professionals in better understanding of man as a being singular that carries experiences, beliefs and culture. However, in many universities these disciplines are taught at the beginning of undergraduate courses and become distant of academic practices, in which the biomedical model is privileged. It is urgent the need to combine knowledge to treat the human being, because the scale of the disease is not restricted to biological sphere. The communication process should be rethought, once I apprehend that the information is given by the health care professional, but it is not understood by the parents. I understand that nursing practice must rethink the care planning or the child with Ostomy, including the father in that context. I reiterate the necessity of nursing involvement in monitoring child and parents and the readjustment of the school by making it more inclusive / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES

O presente estudo buscou compreender os percursos singulares rumo à paternidade no contexto da reprodução assistida, desde a gestação até o primeiro ano de vida do(s) bebê(s). Foi realizado um estudo de caso coletivo longitudinal. Participaram três homens que se submeteram, junto com as esposas, à fertilização in vitro, sendo a causa da infertilidade masculina ou mista. Os pais responderam entrevistas semiestruturadas em três momentos: a) terceiro trimestre de gestação; b) terceiro mês de vida do(s) bebê(s); e c) final do primeiro ano de vida do(s) bebê(s). Os percursos singulares foram relatados e analisados a partir da metodologia do relato clínico. Buscou-se, a partir daí, (re)construir esses três percursos, com seus desafios singulares de elaboração e integração da experiência da paternidade, bem como os desafios no percurso da pesquisadora. Pode-se perceber a solidão dos homens que passaram pela experiência da infertilidade e do tratamento, especialmente por eles terem referido que cuidaram mais do que se sentiram cuidados. Destaca-se a importância de se oferecer um espaço de escuta para esses homens, não só em uma situação de pesquisa, mas também na prática do profissional de saúde mental que trabalha nesse contexto. / This study investigated the unique pathways toward fatherhood in the context of assisted reproduction, from pregnancy to the first year of the babies’ lives. A collective longitudinal case study was conducted. Participants were three men who had undergone, along with their wives, in vitro fertilization. The cause of their infertility was male or mixed. Fathers answered semi-structured interviews in three moments: a) third trimester of pregnancy; b) their babies’ third month of life; c) their babies’ first year of life. The unique pathways have been reported and analyzed based on the clinical report methodology. These three pathways were (re)constructed, with their unique challenges concerning elaboration and integration of fatherhood experience, as well as the researcher’s challenges. Loneliness could be felt in the reports of men who have experienced infertility and treatment. They said they cared more than were cared for. The importance of providing a space for listening to these men is emphasized, not only in a research situation, but also in the practice of mental health professionals who work in this context.

Mudanças recentes na composição e nas relações internas das famílias brasileiras decorreram, entre outros fatores, do número crescente de separações e divórcios e do ingresso maciço das mulheres no mercado de trabalho, de inovações científicas e tecnológicas, como métodos eficazes de contracepção e de reprodução assistida, além de medidas na área da saúde que aumentaram a expectativa de vida da população. O conjunto dessas alterações contribuíram para redefinir a condição feminina, as relações de gênero entre homens e mulheres e as formas de exercício da maternidade e paternidade. Em decorrência dessas transformações, aumentou o número de famílias recompostas constituídas por casais que vivem em segunda união e cuja forma de organização é bastante complexa, sobretudo no que diz respeito às relações com filhos da primeira união dos cônjuges. Em algumas famílias recompostas um homem é pai de filho(s) de união(ões) anterior(es), padrasto do(s) filho(s) da companheira atual e com a qual também pode ter filhos. Esta pesquisa teve por objetivo investigar o exercício da paternidade e as relações conjugais em famílias recompostas constituídas por segunda união de parceiros que têm filho(s) de casamento(s) anterior(es) e eventualmente da segunda união para analisar como homens, que são pais e padrastos, vivenciam a relação de paternidade com filho(s), considerando que aquele(s) da primeira união mora(m) com a ex-esposa, e quais os vínculos que mantêm, enquanto padrasto, com o(s) filho(s) de sua companheira atual. Foram entrevistados sete homens vivendo em segunda união e que são pais e padrastos. Os dados foram coletados com o pai/padrasto mediante entrevista com roteiro semi-estruturado, gravada e transcrita na íntegra, e foram analisados de acordo com a abordagem teórica da antropologia e da psicologia. Os resultados indicam flexibilização das tarefas domésticas e dos cuidados com os filhos, mudanças nas vivências e no exercício da paternidade, procura dos padrastros em manterem boas relações com enteados e com os filhos biológicos com quem não residem. / Recent changes in the composition of Brazilian families were the result of, amongst other factors, the growing number of separations and divorces, the massive entry of women into the market, scientific and technological innovations, like the effective methods of contraception and assisted reproduction as well as measures in the health area that have increased the life expectancy of the population. All these changes contributed to the redefinition of the feminine condition, the gender relations between men and women, and the forms of exercise of motherhood and fatherhood. As a result of these transformations, there was an increase in the number of reconstituted families made up of couples living in second marriage and whose organization is quite complex, especially regarded to relations with children from the first marriage of the spouses. In some recomposed families, a man is the father of previous children, stepfather of the children of the current partner and with whom he may also have children. The objective of this research is to investigate the exercise of fatherhood and conjugal relationships in families who are composed of second union of partners who have children from previous marriage(s) and eventually from second marriage to analyze how men, who are fathers and stepfathers, experience the relationship of fatherhood with children, considering that the one(s) from the first union lives with the ex-wife, and what ties they have as a stepfather of the children of the current companion. Seven men who are fathers and stepfathers and living in second union were interviewed. Data were collected with the father/stepfather through interviews with semi-structured script, recorded and transcribed in its integrity, and analyzed according to the theoretical approaches of anthropology and psychology. The results indicated a flexibilization of the domestic tasks and children caring, changes in the experiences and the exercise of paternity, movement of the stepfathers towards a good relation with the stepchildren, maintenance of the relationship with the consanguineous children.

Este estudo explora, descreve e discute a vivência de luto de cinco adolescentes que perderam o pai biológico por morte de causas diversas. Para tanto, investigaram-se vivências e manifestações relacionadas a essas experiências de perda, suas repercussões para os jovens e para seu desenvolvimento emocional, as estratégias de que eles se valeram para enfrentar esse luto, sua resiliência frente à perda vivida (capacidade do jovem de resistir às adversidades geradas pela morte do pai, adaptar-se e manter-se psicologicamente saudável), identificando-se fatores de risco (vulnerabilidades) e de proteção para a elaboração desse luto e o desenvolvimento dos adolescentes, bem como a necessidade ou não de se proporem formas de cuidado específicas para os enlutados, entre as inúmeras possibilidades terapêuticas, mesmo que profiláticas. Como procedimento, usaram-se o Inventário de Auto-avaliação para jovens de 11 a 18 anos (YSR), para rastrear a eventual presença de problemas de saúde mental, e uma entrevista semi-estruturada com roteiro. A análise dos dados obtidos estabeleceu que o luto pela morte do pai biológico na adolescência tem características próprias, distinguindo-se do luto que essa perda acarreta em outras etapas do ciclo vital, em função dos desafios singulares do desenvolvimento nesse período. Embora a morte do pai biológico represente para o adolescente uma crise, também pode ser uma oportunidade para que, no enfrentamento dessa perda, ele amadureça. Constatou-se ainda a importância do suporte social de uma rede de apoio que compreenda e atenda as necessidades do enlutado, sobretudo pela presença de uma mãe funcional, para a ressignificação dessa perda, além de que, se o adolescente tem dificuldade para expressar seu luto, pode ficar entorpecido e negar as conseqüências dessa perda, comprometendo seu desenvolvimento emocional. / This study explores, describes and discuss the bereavement experience of five adolescents who had lost their biological father by death of several causes. Thus, it was sought to investigate the experiences and events related to these experiences of loss; its impact on young people and for their emotional development; the strategies that they used to cope with this mourning; the resilience to face the loss experienced (the adolescents ability to resist to the adversities generated by the death of the father, adapt to and remain psychologically healthy), identifying risk factors (vulnerabilities) and protection factors for the resolution of this bereavement and the development of adolescents, and if its necessary to propose ways of special care for the bereaved, among the many therapeutic possibilities, even if prophylactic. As procedure, were used the Inventory of Self-assessment for young people from 11 to 18 years-old (YSR), to trace the possible presence of mental health problems and a semistructured interview with a script. The analysis of data obtained established that the mourning for the death of biological father in adolescence presents its own characteristics, distinguishing itself from the grief that this loss brings at other times of the life cycle, according to the natural challenges of development in that period; the fact that, although the death of the biological father represents a crisis for the adolescent, can also be an opportunity for him or her, when facing this loss, matures; The importance of social support of a supportive network that understands and meets the needs of the bereaved, especially the presence of a functional mother, for the re-signification of this loss, as well as the fact that the teenagers difficulties to express themselves about the mourning that they experience can guide them to remain numb and deny the consequences of this loss, and impair their emotional development.

Summary: In order that one may approach the main theme of this thesis, the massacre at Poverty Bay, the events leading there to, and the effects thereof, some slight knowledge is required of the general situation in New Zealand in the years immediately preceding. The writer considers that no apology is needed for introducing into a work that has for its title "The Poverty Bay Massacre of 1868", such apparently unrelated topics as the campaigns of 1845 to 1868 and the cult of Pai-Marire. In any historical review, cause and effect are so closely related that it is difficult, when choosing a particular field of research, to decide how much to include and, more important, how much to discard. The Maori Wars in the Waikato and Taranaki districts spread through the whole of the North Island and in time to the East Coast. Here they were intensified by the blend of Christianity and savage barbarity that is known as Pai-Marire or Hauhauism. It was during this campaign on the Coast that Te Kooti Rikirangi first came into political prominence. A study of Hauhauism is interesting in itself; reviewed in connection with the central figure of this thesis, Te Kooti, it acquires new significance. It is doubtful how far he was sincere in the religious ritual he instituted, itself a modification, and an adaptation of the old Pai-Marire cult, but out of it he fashioned a powerful weapon against the Pakeha. Perhaps it deserves a place as a psychological study of the influence of community worship in a mysterious and militant ritual upon the religious emotionalism of any sect and when, as here, that sect was composed of semi-civilised or wholly savage Maoris at a critical period in the history of their race, it was inevitable that it should issue in action. This thesis represents an attempt at more than merely collecting loose threads about the massacre into one narrative. It has been the writer�s aim to place the massacre in its right setting in the history of New Zealand, and thus to show its significance. The method chosen has been that of grouping the events round one central figure, that of the perpetrator of the massacre, and the first four chapters therefore are incidental though necessary. The history of this man continues long after 1868, the date of the massacre and a small section has been added to cover the period 1868 to 1893, the year of his death on the plea that though irrelevant to the subject of the thesis it gives a rounded effect that might otherwise be lacking--Introduction.

<p>Inhibitors belonging to the serpin (serine protease inhibitor) family control proteases involved in various physiological processes. All serpins have a common tertiary structure based on the dominant b-sheet A, but they have different inhibitory specificity. The specificity of a serpin is determined by the Pl-Pl’ peptide bond acting as a bait for the target protease which is made up of an exposed reactive centre loop (RCL). The serpin plasminogen activator inhibitor type-1 (PAI-1) is the main physiological inhibitor of urokinase-type and tissue-type plasminogen activators (uPA and tPA, respectively). Elevated plasma levels of PAI-l have been correlated with a higher risk of deep venous thrombosis, and PAI-1 is a risk factor for recurrent myocardial infarction. Furthermore, PAI-1 has a role in cell migration and has been suggested to regulate tumor growth and angiogenesis. PAI-1 is unique among the serpins in that it can spontaneously and rapidly convert into its latent form. This involves full insertion of the RCL into b-sheet A. </p><p>There were two partially overlapping goals for this thesis. The first was to use latent PAI-1 as model for development of a fluorescence-based method, Donor-Donor Energy Migration for intramolecular distance measurements. The second goal was to use DDEM, together with other biochemical methods, to reveal the structure of the PAI-1/uPA complex, the conformation of the RCL in active PAI-1, and molecular determinants responsible for the conversion of PAI-1 from the active to the latent form.</p><p>The use of molecular genetics for introduction of fluorescent molecules enables the use of DDEM to determine intramolecular distances in a variety of proteins. This approach can be applied to examin the overall molecular dimensions of proteins and to investigate structural changes upon interactions with specific target molecules. In this work, the accuracy of the DDEM method has been evaluated by experiments with the latent PAI-1 for which X-ray structure is known. Our data show that distances approximating the Förster radius (57±1 Å) obtained by DDEM are in good agreement (within 5.5 Å) with the distances obtained by X-ray crystallography.</p><p>The molecular details of the inhibitory mechanism of serpins and the structure of the serpin/protease complex have remained unclear. To obtain the structural insights required to discriminate between different models of serpin inhibition, we used fluorescence spectroscopy and cross-linking techniques to map sites of PAI-1/uPA interaction, and distance measurement by DDEM to triangulate the position of the uPA in the complex. The data have demonstrated clearly that in the covalent PAI-1/uPA complex, the uPA is located at the distal end of the PAI-1 molecule relative to the initial docking site. This indicates that serpin inhibition involves reactive center cleavage followed by full loop insertion, whereby the covalently linked protease is translocated from one pole of the inhibitor to the opposite one. </p><p>To search for molecular determinants that could be responsible for conversion of PAI-1 to the latent form, we studied the conformation of the RCL in active PAI-1 in solution. Intramolecular distance measurements by DDEM, the newly a developed method based on probe quenching and biochemical methods revealed that the RCL in PAI-1 is located much closer to the core of PAI-1 than has been suggested by the recently resolved X-ray structures of stable PAI-1 mutants, and it can be partially inserted. This possibly explains for the ability of PAI-1 to convert spontaneously to its latent form. </p>

<p>In this thesis, different aspects of the insulin resistance syndrome in relation to plasminogen activator inhibitor-1 (PAI-1) activity are investigated in a longitudinal population-based study. Participants were men investigated at ages 50 and 70 with follow-up data on mortality.</p><p>High PAI-1 activity was associated with low insulin sensitivity, high concentrations of serum triglycerides, high body mass index and high waist/hip ratio, independently of each other and of potential confounders. Low birth weight predicted high blood pressure, insulin resistance, truncal obesity and high PAI-1 activity but not the abdominal obesity or dyslipidaemia present in the insulin resistance syndrome. Increased physical activity level between 50 and 70 years of age, in the absence of active intervention, was associated with improved glucose, insulin, proinsulin and lipoprotein metabolism. Insulin and proinsulin seemed to be important factors that mediate much of the association between a sedentary lifestyle and increased risk of cardiovascular disease. The reported dietary intake of both mono- and polyunsaturated fatty acids was positively associated with PAI-1 activity, whereas saturated fatty acid intake displayed no association. The associations present between PAI-1 activity and the fatty acid proportions in serum cholesterol esters were partly influenced by factors related with the insulin resistance syndrome.</p><p>This thesis provides further knowledge to the epidemiological view of the interrelations of the insulin resistance syndrome, PAI-1, birth weight, and lifestyle factors as physical activity and dietary habits. PAI-1 is a part of the insulin resistance syndrome and is associated both with modifiable and non-modifiable factors related with this syndrome.</p>

Des données antérieures obtenues dans notre laboratoire ont montré le rôle essentiel de PAI-1 dans la croissance de cellules cancéreuses dorigine murine et linduction dune angiogenèse concomitante (Bajou et al, 1998;Bajou et al, 2001). Bien que ces résultats aient été confirmés par dautres groupes dans différents modèles expérimentaux (Gutierrez et al, 2000;McMahon et al, 2001), très peu dinformations concernant limplication de PAI-1 dans la progression des carcinomes humains étaient disponibles dans la littérature. Cest pourquoi, dans le premier chapitre de ce travail, nous nous sommes intéressés au comportement de différentes lignées de carcinomes humains lorsquelles sont placées dans un environnement dépourvu en PAI-1. Lensemble des résultats obtenus est décrit dans la première publication intitulée : « Host plasminogen activator inhibitor-1 promotes human skin carcinoma progression in a stage-dependent manner » (Maillard et al, 2005, Neoplasia, 7: 57-66) La synthèse des différentes études utilisant différents modèles expérimentaux in vivo (voir introduction, Tableau 6) a montré quelques divergences quant à linfluence de PAI-1 dans la progression tumorale et la formation de métastases. Ces résultats contradictoires pourraient sexpliquer en partie par la grande diversité des modèles expérimentaux utilisés. Les lignées de cellules tumorales utilisées, le nombre de cellules implantées, ainsi que le site dimplantation chez les animaux sont quelques uns des facteurs pouvant influencer les résultats. Dans ce contexte, lutilisation danimaux transgéniques développant spontanément des lésions tumorales est une alternative attrayante pour étudier limpact dun facteur sur lévolution de ces lésions. A lheure actuelle, un seul groupe sest intéressé à limpact dun déficit en PAI-1 dans un modèle de souris développant de manière spontanée des adénocarcinomes mammaires qui métastasent dans les poumons, et na pas observé de différences entre les animaux déficients en PAI-1 et leurs contrôles de type sauvage (Almholt et al, 2003). Ce deuxième chapitre est consacré à létude de limplication de PAI-1 dans un autre modèle murin de cancérogenèse spontanée, à savoir, les souris TRP-1/SV40 Tag caractérisées par un développement de tumeurs oculaires qui forment des métastases dans le cerveau. Ce travail a été réalisé en collaboration avec le Laboratoire de Vectorologie et de Transfert de Gènes (Docteur M. Perricaudet, UMR8121, Villejuif, France), au cours dun séjour de plus de 6 mois. Lensemble des résultats obtenus est décrit dans la seconde publication intitulée : « Reduction of brain metastases in transgenic ocular tumor in plasminogen activator inhibitor-1 deficient mice » (Maillard et al, soumis pour publication) Les résultats présentés dans le troisième chapitre mettent en avant lintérêt des chambres de transplantation in vivo de kératinocytes tumoraux comme modèle détude des interactions hôte-tumeur. Dans un premier temps, ce modèle nous a permis de déterminer limportance de lorigine cellulaire de PAI-1 (cellules tumorales ou cellules hôtes) sur son effet pro-angiogène. Dautre part, des effets opposés de PAI-1 en fonction de sa concentration ont été mis en évidence et apportent des éléments de réponse quant aux effets pro- ou anti-angiogènes de cet inhibiteur des activateurs du plasminogène. Les résultats sont repris dans la troisième publication intitulée : « Host-derived plasminogen activator inhibitor-1 (PAI-1) concentration is critical for in vivo tumoral angiogenesis and growth » (Bajou et al, 2004, Oncogene, 23:6986-6990) Dans un second temps, en collaboration avec léquipe du Professeur J. Boniver (Laboratoire dAnatomie et de Cytologie Pathologiques, ULg, Liège), les chambres de transplantation ont été utilisées pour évaluer in vivo lefficacité thérapeutique du GM-CSF comme cytokine favorisant la migration intratumorale de cellules dendritiques matures. Ces travaux sont présentés dans la quatrième publication intitulée : « Delivery of granulocyte-macrophage colony-stimulating factor in bioadhesive hydrogel stimulates migration of dendritic cells in models of human papillomavirus-associated (pre)neoplastic epithelial lesions » (Hubert et al, 2004, Antimicrobial agents and chemotherapy, 48: 4342-4348)

In this thesis, different aspects of the insulin resistance syndrome in relation to plasminogen activator inhibitor-1 (PAI-1) activity are investigated in a longitudinal population-based study. Participants were men investigated at ages 50 and 70 with follow-up data on mortality. High PAI-1 activity was associated with low insulin sensitivity, high concentrations of serum triglycerides, high body mass index and high waist/hip ratio, independently of each other and of potential confounders. Low birth weight predicted high blood pressure, insulin resistance, truncal obesity and high PAI-1 activity but not the abdominal obesity or dyslipidaemia present in the insulin resistance syndrome. Increased physical activity level between 50 and 70 years of age, in the absence of active intervention, was associated with improved glucose, insulin, proinsulin and lipoprotein metabolism. Insulin and proinsulin seemed to be important factors that mediate much of the association between a sedentary lifestyle and increased risk of cardiovascular disease. The reported dietary intake of both mono- and polyunsaturated fatty acids was positively associated with PAI-1 activity, whereas saturated fatty acid intake displayed no association. The associations present between PAI-1 activity and the fatty acid proportions in serum cholesterol esters were partly influenced by factors related with the insulin resistance syndrome. This thesis provides further knowledge to the epidemiological view of the interrelations of the insulin resistance syndrome, PAI-1, birth weight, and lifestyle factors as physical activity and dietary habits. PAI-1 is a part of the insulin resistance syndrome and is associated both with modifiable and non-modifiable factors related with this syndrome.

<p>The development of improved sequencing technologies hasenabled the field of genomics to evolve. Handling andsequencing of large numbers of samples require an increasedlevel of automation in order to obtain high throughput andconsistent quality. Improved performance has lead to thesequencing of numerous microbial genomes and a few genomes fromhigher eukaryotes and the benefits of comparing sequences bothwithin and between species are now becoming apparent. Thisthesis describes both the development of automated purificationmethods for DNA, mainly sequencing products, and a comparativesequencing project.</p><p>The initially developed purification technique is dedicatedto single stranded DNA containing vector specific sequences,exemplified by sequencing products. Specific capture probescoupled to paramagnetic beads together with stabilizing modularprobes hybridize to the single stranded target. After washing,the purified DNA can be released using water. When sequencingproducts are purified they can be directly loaded onto acapillary sequencer after elution. Since this approach isspecific it can be applied to multiplex sequencing products.Different probe sets are used for each sequencing product andthe purifications are performed iteratively.</p><p>The second purification approach, which can be applied to anumber of different targets, involves biotinylated PCR productsor sequencing products that are captured using streptavidinbeads. This has been described previously, buthere theinteraction between streptavidin and biotin can be disruptedwithout denaturing the streptavidin, enabling the re-use of thebeads. The relatively mild elution conditions also enable therelease of sensitive biotinylated molecules.</p><p>Another project described in this thesis is the comparativesequencing of the 40 kb<i>cag</i>pathogenicity island (PAI) in four<i>Helicobacter pylori</i>strains. The results included thediscovery of a novel gene, present in approximately half of theSwedish strains tested. In addition, one of the strainscontained a major rearrangement dividing the<i>cag</i>PAI into two parts. Further, information about thevariability of different genes could be obtained.</p><p><b>Keywords:</b>DNA sequencing, DNA purification, automation,solid-phase, streptavidin, biotin, modular probes,<i>Helicobacter pylori</i>,<i>cag</i>PAI.</p>