Der Prozess der Entzündung und die Entstehung von Neoplasien sind durch komplexe Abläufe innerhalb des Organismus gekennzeichnet, die einer angemessenen Kontrolle des Zellzyklus und der Proliferation bedürfen. Dabei spielt eine Reihe von Eiweißstoffen, sowohl im Entzündungsgeschehen als auch in der Entstehung von Malignomen, eine wesentliche Rolle. Das proinflammatorische Zytokin Macrophage Migration Inhibitory Factor (MIF) ist eines dieser Proteine, welches einen bedeutenden Beitrag in der Proliferation und Differenzierung von Zellen leistet. MIF stimuliert die Tumorzellproliferation, die Migration und die Metastasierung, fördert die Tumorangiogenese und hemmt die p53-vermittelte Apoptose. Aufgrund einer vermehrten Expression von MIF in diversen malignen Tumoren interessierte das Vorkommen von MIF im Zervixkarzinom. In der vorliegenden Arbeit wurde der Expressionsstatus von MIF im Zervixkarzinom auf mRNA- und Proteinebene mittels Western Blot und Immunhistochemie untersucht. Zum Nachweis der mRNA Expression von MIF im Zervixkarzinom wurde RNA aus drei gesunden Gewebeproben der Zervix, aus vier Plattenepithelkarzinomen und aus einem Adenokarzinom der Zervix isoliert. Dabei zeigte sich eine signifikante Überexpression der MIF mRNA im Zervixkarzinom gegenüber dem gesunden Zervixgewebe. Des Weiteren konnte im Rahmen einer Western Blot Analyse ebenfalls eine Überexpression von MIF Protein in zervikalen Tumorzellen, SiHa und CaSki, nachgewiesen werden. Außerdem zeigte sich auch immunhistochemisch eine MIF Überexpression. Dabei wurden Paraffinschnitte von Zervixkarzinomen (n = 25), Carcinomata in situ und CIN III (n = 23) und Zervixdysplasien (n = 32) immunhistochemisch gefärbt. Da beim Zervixkarzinom besonders die Radiotherapie meist in Kombination mit einer Chemotherapie von zentraler Bedeutung ist, rückte die Frage nach einer möglichen Modulation der Strahlensensitivität durch MIF in das klinische Interesse. Daher wurden Zervixkarzinomzellen mit einer Strahlendosis von 2,5 bis 15 Gy in An- und Abwesenheit eines MIF-Inhibitors bestrahlt und anschließend die Bestrahlungszytotoxizität dieser Zellen mithilfe eines Klonogenitätsassays näher beleuchtet. Mit steigender Strahlendosis nahmen die überlebenden zervikalen Tumorzellen in Abhängigkeit vom MIF-Inhibitor ab. Eine mögliche Bedeutung von MIF bzw. des MIF-Inhibitors, sowohl als Radiosensitizer im Rahmen der Strahlentherapie des Zervixkarzinoms als auch als Screeningparameter, bedarf noch weiterer Studien.

I dagsläget kan WM-datas Fältmodul i deras MoveITS-system inte hantera kartdata i andra format än Shape. Fältmodulen är en TabletPC som kör operativsystemet Windows XP. Den kan användas för att redigera viss geografisk information som exempelvis skyltpositioner. Fältmodulen används av Stockholms Tekniska kontor för inventering av skyltstolpar.Stockholms Tekniska kontor ska börja leverera sina kartor i GML (Geography Markup Language). Men eftersom WM-datas Fältmodul inte klarar av det formatet skulle det här examensarbetet gå ut på att ta fram komponenter för hantering av det. Då det under examensarbetets gång har varit svårt att få tag i information runt GML har istället en större fokus blivit lagd på MIF (MapInfo Interchange Format). Eftersom det finns andra kommuner som använder MIF finns det intresse från WM-data att det tas fram komponenter även för detta format.Ett stort antal klasser har utvecklats för hantering av MIF-filer. Dessa klasser är helt utvecklade i C# och har gjorts under examensarbetets gång utifrån de specifikationer som finns för formatet från företaget MapInfo. För GML har det tagits fram information som kan ligga till grund för utveckling av komponenter för hantering av det formatet.

Les sepsis sévères et chocs septiques constituent des causes croissantes de morbidité et de mortalité chez les patients hospitalisés. La dysfonction cardio-circulatoire lors d'un choc septique s'avère un élément majeur et initial dans l'induction et l'entretien des défaillances organiques subséquentes. En effet, certaines bactéries et endotoxines déclenchent une séquence d'événements cellulaires qui mènent à une altération des performances cardiaques. Le macrophage migration inhibitory factor (MIF) a récemment été identifié en tant que facteur de dépression cardiaque lors de choc septique. Ainsi, la présente étude révèle que la neutralisation du MIF prévient la dysfonction myocardique induite par l'endotoxine dans un modèle expérimental chez le rat. De façon sous-jacente, le blocage du MIF empêche la hausse d'expression et de production dans le coeur de cytokines inflammatoires paracrines et autocrines (IL-1, IL-6 et TNF-[alpha]) en réponse au LPS. D'un point de vue mécanistique, cette étude démontre que la neutralisation du MIF inhibe l'apoptose des cardiomyocytes provoquée par l'endotoxine. Ainsi, le présent travail met en évidence un mécanisme précis par lequel le MIF influence la dépression cardiaque endotoxinique. Ce mécanisme implique que le blocage du MIF induit une augmentation du ratio protéique cardiaque de Bcl-2/Bax contribuant ainsi à prévenir le relâchement du cytochrome c mitochondrial induit par l'endotoxine. Cette inhibition de la perte de cytochrome c mitochondrial entraîne une réduction de l'activation de la caspase-3. La neutralisation du MIF rétablie la déficience provoquée par le LPS au niveau de la translocation nucléaire de phospho-Akt et par conséquent l'expression du facteur nucléaire de survie cardiaque GATA-4. Cette baisse d'activité caspase-3 et le rétablissement de la translocation/expression des facteurs de survie par le blocage du MIF résultent en une diminution de la fragmentation de l'ADN caractéristique de l'apoptose tardive. Globalement, l'inactivation du MIF lors d'un choc endotoxinique prévient le déséquilibre inflammatoire et apoptotique dans le coeur protégeant ainsi contre la dysfonction myocardique subséquente.

The complex process of wound repair becomes disrupted in the elderly with a profound effect on patient morbidity and huge financial implications for the NHS. While age itself is a risk factor for delayed healing recent work implicates estrogen decline, rather than intrinsic ageing per se, as the critical regulator of delayed healing in elderly subjects. In women estrogen levels fall dramatically post-menopause and with increasing life expectancy most women in the developed now world spend at least a third of their lives in a state of estrogen deprivation. Estrogen replacement can reverse this delay, but unfortunately long term estrogen treatment (HRT) increases breast cancer risk such that steroidal estrogen is now listed as a carcinogen. The aim of this study has been to functionally dissect the role of estrogen signalling during repair at the molecular, cellular and physiological levels. New data presented within this thesis reveal estrogen to be a global regulator of healing with pleiotropic effects on multiple wound cell types. By combining pharmacological manipulation and genetic ablation my data reveals novel diametrically opposed roles for the two estrogen receptor isoforms, ERalpha and ERbeta, during healing. I have further exploited this to demonstrate the in vivo therapeutic potential of compounds with receptor selective agonistic/antagonistic activity. Additionally, I have further investigated the mechanism of action of estrogen and these selective compounds implicating the pro-inflammatory cytokine macrophage migration inhibitory factor (MIF) in beneficial effects on healing. This research leads the way toward translation into human studies with the ultimate aim of developing targeted therapeutics for the treatment of delayed acute and chronic wounds, particularly in the elderly.

Macrophage migration inhibitory factor (MIF) has been implicated in mediating both innate and adaptive immune responses in inflammatory and infectious diseases. The sequence and structure of MIF is highly conserved across the avian phylogeny, which underlies high sequence homology and functional similarities between turkey and chicken MIFs. Turkey MIF (TkMIF) inhibited cell migration and promoted cell proliferation with production of inflammatory mediators, comparable to the biological properties of chicken MIF (ChMIF), thus indicating the biological cross-reactivity between turkey and chicken MIFs. This study identified the cell surface receptor(s) that could bind ChMIF and the biological roles triggered by such interactions. In addition to CD74, a previously identified receptor, CXCR4 also interacts with ChMIF. Moreover, the formation of receptor complexes was shown between CXCR4 and CD74. MIF signaling through CXCR4 and CD74 led to cell chemotaxis and proliferation activity as well as intracellular calcium influx. Intriguingly, Eimeria MIF (EMIF), a homologue secreted following parasitic infection, also interacted with CD74 leading to comparable biological functions to those of ChMIF. Given such observations, we hypothesized that CXCR4 and CD74 are receptors for ChMIF leading to the functional consequences similarly manifested by EMIF interaction with the corresponding receptors. EMIF, predominantly secreted from the invasive merozoite stage, may help the parasite exploit the host immune response by interacting with common ChMIF receptors. This may lead to functional mimicry thus provoking the question of whether EMIF would modulate the biological functions of ChMIF to manipulate the host defense that allows more efficient invasion of the host. To evaluate this concept, a transgenic E. tenella lacking MIF was generated by in vivo passage of E. tenella transfected with a CRISPR plasmid targeting EMIF. Although not fully disrupted, reduction of EMIF expression was observed in the transgenic E. tenella itself as well as in inoculated cells, which resulted in enhanced survival of host cells. Herein, we achieved a better characterization of the functional roles of both avian and parasite MIFs underlying the interaction with common host receptors, along with the essential role of parasite MIF promoting host cell death during parasitic infection. / PHD / When animals get infected or injured, their immune system senses invading pathogens or damaged tissues as danger signals, which often elicits the production of inflammatory mediators. These are chemical messengers secreted mostly by immune cells that initiate cellular communication and infiltration of immune cells to the infection/damaged site leading to inflammatory responses to eliminate the infectious agents and repair damaged tissues. Among many inflammatory mediators, macrophage migration inhibitory factor (MIF) is involved in inflammatory and immune response by regulating cell migration. Interestingly, MIF is secreted by Eimeria parasites (that cause the costly coccidiosis disease in poultry) as well as by chickens (host animal) after infection with this pathogen. Toward a better understanding of the impacts of both avian and parasite MIFs on the host immune response, three specific studies were completed. First, MIF displayed high degree of gene sequence identity and functional similarity between chicken and turkey, supporting the evolutionarily conservation of MIF across birds. The second study identified the MIF receptors and their complexes, which engage in the biological functions of chicken MIF. Through binding to these cell surface receptors, chicken MIF can regulate cell migration and proliferation with calcium release. Intriguingly, Eimeria MIF secreted after parasitic infection is able to bind the same receptors leading to comparable biological functions to those of chicken MIF. Lastly, the role of Eimeria MIF was further evaluated by disrupting its gene in the parasite. Although not fully disrupted in the transgenic parasites, its expression was decreased resulting in enhanced survival of host cells, thus suggesting a deleterious effect of Eimeria MIF on the host, as well as its potential as a therapeutic target to control coccidiosis in poultry.

Inflammation can be initiated by an innate immune sensor, followed by activation of a signal mediator, resulting in control of immune response by a signal regulator. Mammalian nucleotide-binding oligomerization domain protein 1 (Nod1) and Nod2 initiate host innate immune response by recognition of specific bacterial molecules, resulting in the production of pro-inflammatory cytokines, chemokines, and anti-microbial peptides. A candidate sequence of chicken Nod1 (ChNod1) was identified with no current evidence of ChNod2. Stimulation of transiently overexpressed ChNod1 and its mutants with mammalian Nod-specific ligands was not conclusive of the function of ChNod1 most likely due to self-activation of ChNod1. In vitro studies showed no significant difference in expression of Nod1, its signaling molecules and pro-inflammatory cytokines in stimulated chicken mononuclear cells with synthetic ligands for mammalian Nod1 or Nod2. A signal mediator, macrophage migration inhibitory factor (MIF) inhibits the random migration of macrophages. Chemotaxis assay using recombinant ChMIF (rChMIF) revealed a substantial decrease in migration of macrophages. qRT-PCR analysis revealed that the presence of rChMIF enhanced levels of IL-1β and iNOS during monocytes stimulation with LPS. Additionally, Con A-stimulated lymphocytes exhibited enhanced IFN-γ and IL-2 transcripts in the presence of rChMIF. IL-22, which may act as a signal regulator, is an important effector of activated Th1 and Th17 as well as natural killer cells during inflammation. Recombinant ChIL-22 alone did not have an impact on chicken embryo kidney epithelial cells (CKECs); however, co-stimulation of CKECs with LPS and rChIL-22 enhanced the production of pro-inflammatory cytokines and anti-microbial peptides. Furthermore, rChIL-22 alone stimulated acute phase reactants in chicken embryo liver cells. These effects of rChIL-22 were abolished by addition of rChIL22 binding protein. Taken together, these results indicate an important role of ChIL-22 on epithelial cells and hepatocytes during inflammation. In this project, we identified and characterized the avian inflammatory mediators ChNod1, ChMIF, and ChIL-22. Studying each of their biological function in avian inflammation, especially under pathogenic challenges in epithelial tissues will provide a foundation for understanding the role of these inflammatory mediators in mucosal immunity. / Ph. D.

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Made available in DSpace on 2014-06-11T19:30:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-12-19Bitstream added on 2014-06-13T20:00:29Z : No. of bitstreams: 1 borges_jbc_dr_botfm.pdf: 523209 bytes, checksum: 91e394791a292a14faa66bdc26898c97 (MD5) / Os serviços de saúde acompanhando o desenvolvimento econômico e alta competitividade direcionam atenção à qualidade com que os serviços estão sendo realizados. A avaliação de um serviço de saúde está ligada à percepção do paciente na medida em que suas expectativas foram atendidas na efetividade e na qualidade das intervenções. O objetivo deste estudo consistiu em avaliar a funcionalidade e a qualidade de serviço em pacientes de cirurgia cardíaca no período pré-operatório e pós-operatório recente 6ºPO. Buscou-se ainda relacionar o nível de funcionalidade e de qualidade do serviço com: gênero, faixa etária e uso de circulação extra-corpórea (CEC). Foram estudados 82 pacientes, submetidos à cirurgia cardíaca eletiva, operados por toracotomia médio esternal, de ambos os gêneros, com idade entre 31 e 83 anos. Os pacientes entraram no estudo consecutivamente, de acordo com os critérios de inclusão e exclusão, a partir de março a setembro de 2006. Os pacientes foram divididos em subgrupos de acordo com: gênero, faixa etária, tipo de cirurgia, primeira cirurgia cardíaca ou reoperação e uso de CEC. O nível de funcionalidade foi avaliado pela escala MIF e a qualidade dos serviços prestados pela escala SERVQUAL-Card, escala modificada da SERVQUAL. A funcionalidade foi prejudicada após a cirurgia cardíaca até o 6º PO. Os níveis mais elevados de perda funcional ocorreram na categoria de locomoção, concentrando-se na atividade de subir escadas. No pré-operatório os pacientes mais jovens apresentaram níveis mais elevados de funcionalidade. Os níveis diminuíram com o aumento da idade dos pacientes e a menor funcionalidade foi na faixa etária de 70 a 85 anos. Tanto no aspecto motor como no cognitivo a utilização de CEC não esteve relacionada com o grau de perda funcional no pós-operatório recente. A qualidade dos serviços percebida pelo paciente foi satisfatória. / The health services, following the economical development and the high competition, focus its attention on the quality of services. The evaluation of a health service is related to the patient perception as his expectations are met in the quality and effectiveness of the interventions.This paper was aimed at evaluating the service functionality and quality in cardiac surgery patients during the preoperative and recent postoperative periods (6° PO). It was also aimed at relating the service level of functionality and quality with the following: gender, age, and use of extra-corporal circulation (ECC). Eight two patients (men and women), subjected to elective cardiac surgery, were studied; they were operated by means of sternal medium thoracotomy and their ages ranged from 31 to 83 years. The patients were entered the study consecutively, according to the criteria of inclusion and exclusion, from March to September, 2006. The patients were divided into sub-groups in accordance with gender, age, type of surgery, first cardiac surgery or re-operation and use of ECC. The functionality level was assessed by using the MIF scale and the quality of the services was measured by the SERVQUAL scale-Card, modified SERQUAL scale. The functionality was harmed after the cardiac surgery until the 6° PO. The most elevated levels of functional losses occurred in the locomotion category, concentrating on the activity of climbing stairs. During the preoperative period, the youngest patients presented more elevated levels of functionality. The levels were decreased according to the increase in the patients ages and the smallest functionality was found in the ages of 70 to 85 years. Both in the motor aspect and in the cognitive aspect, the use of ECC was not related to the degree of functional loss during the recent postoperative period. The quality of the services watched by the patients was considered satisfactory.

Les pucerons évitent ou contrôlent les défenses de leur plante hôte par des mécanismes moléculaires qui sont actuellement peu élucidés / Aphids attack virtually all plant species and cause serious crop damages in agriculture