INTRODUÇÃO: O ozônio (O3) é encontrado na natureza e também pode ser produzido no corpo humano através da ativação de anticorpos. Seus efeitos anti-bactericidas são descritos na literatura, mas esses dados são controversos quanto a um potencial efeito benéfico da ozonioterapia no tratamento de certos tipos de infecção. OBJETIVO: Avaliar os efeitos da aplicação intraperitoneal (i.p.) de uma mistura gasosa de ozônio em um modelo de ligadura e punção de ceco (LPC) em ratos, através da dosagem de interleucinas (IL)-6, IL-10 e da quimiocina CINC-1 (cytokine-induced neutrophil chemoattractant), da lesão pulmonar aguda (LPA) e da análise das taxas de sobrevida. MÉTODO: Quatro grupos de ratos Wistar foram utilizados para análise de cada objetivo (CTR, LPC, LPC+O2 e LPC+O3). Os animais do grupo CTR foram submetidos somente a laparotomia. O grupo LPC foi submetido aos procedimentos de LPC. Os outros grupos foram submetidos à LPC e receberam injeção (i.p.) da mistura gasosa correspondente, administrada a cada 12 horas durante o período de observação. Os níveis séricos de IL-6, CINC-1 e IL-10 foram determinados por imuno- ensaio (enzyme linked immunosorbent assay- ELISA). A LPA foi avaliada através da histologia pulmonar e quantificada através do método do extravasamento pulmonar do Azul de Evans. Os animais da análise de sobrevivência foram observados por cinco dias. Os valores obtidos foramexpressos como médias ± erro-padrão da média (EP) ou medianas mais percentis 25 e 75(P25; P75), de acordo com a distribuição dos dados. Considerou-se significante p<0,05. RESULTADOS: Os ratos do grupo CTR exibiram os menores níveis de CINC-1 (p<0,01). O grupo LPC+O3 teve níveis menores de CINC-1 comparado a LPC+O2 e LPC (p<0,05). Os níveis de IL-10 do grupo CTR foram menores do que nos outros 3 grupos(p=0,02) . Não houve diferenças entre os outros 3 grupos (p=0,85). IL-6 foi significativamente menor para o grupo CTR (30,8± 4,8) quando comparado a todos os outros grupos (p<0,001). LPC+O3 e LPC+O2 exibiram níveis menores quando comparados ao grupo LPC (p<0,01). Não houve diferença entre os grupos LPC+O3 e LPC+O2 (p=0,54). O escore de histologia pulmonar foi menor para CTR (p=0,02). Os outros grupos não apresentaram diferenças significantes intergrupos (p=0,3). Os valores dos coeficientes de extravasamento pulmonar do Azul de Evans foram menores para LPC+O3 quando comparado aos grupos LPC+O2 e LPC (p=0,02), porém não houve diferença na comparação com CTR O grupo CTR teve o maior tempo de sobrevida (110±10h) comparado com os outros grupos, ou seja, LPC (57,3± 10,4h), LPC+O2 (71 ± 12,9h) e LPC+O3 (52,1 ± 8), os quais não apresentaram diferenças entre si quanto à sobrevida (p=0,4). CONCLUSÃO: No presente estudo experimental em ratos, a ozonioterapia teve um benefício potencial na modulação da resposta inflamatória e na LPA, mas não influenciou as taxas de sobrevida dos animais. / INTRODUCTION: Ozone (O3) is found in nature and also can be produced in the human body through activation of antibodies. Its antibacterial effect has been described in the literature, but these data are controversial regardi ng a benefic role of O3 therapy in the treatment of certain types of infection. OBJECTIVE: To evaluate the effects of intraperitoneal (i.p.) application of an O3 gas mixture in a rat model of cecal ligation and puncture (CLP), by analyzing interleukin (IL)-6, IL-10 and cytokine-induced neutrophil chemoattractant (CINC)-1 levels, acute lung injury (ALI) and survival rates. METHOD: Four animal groups were used (SHAM, CLP, CLP+O2 and CLP+O3). SHAM animals were submitted solely to laparotomy. CLP group was submitted to cecal ligation and puncture. The other groups were submitted to CLP and received injections (i.p.) of the corresponding gas mixture every 12 hours during the observation period. The serum concentrations of IL-6, CINC-1 and IL-10 were determined by the enzyme-linked immunosorbent assay (ELISA). ALI was evaluated with pulmonary histology and quantitated by means of the Evans blue dye (EBD) lung leakage method. For survival analysis, animals were observed for 5 days. Values were expressed as means ± SEM or medians (P25; P75), according to the data distribution. A p<0,05 was considered significant.RESULTS: SHAM rats had the lowest levels of CINC-1 compared to all other groups (p<0,01). CLP+O3 group had lower levels of CINC-1 compared to CLP+O2 and CLP (p<0,05). SHAM IL-10 levels were the lowest compared to the 3 other groups (p=0,02). There were no differences between the other 3 groups (p=0,85). IL-6 was significantly lower for SHAM compared to all groups (p<0,001). CLP+O3 and CLP+O2 had lower levels when compared to CLP (p<0,01). Comparison between groups CLP+O3 and CLP+O2 showed no significant difference (p=0,54). Pulmonary histology score was lower for SHAM (p=0,02). The other groups presented no statistical difference when compared to each other (p=0,3). EBD lung leakage values were lower to CLP+O3 compared to CLP+O2 and CLP (p=0,02). SHAM group had the longest survival time (110±10h) compared to all other groups (p=0,002). CLP (57,3± 10,4h), CLP+O2 (71 ± 12,9h) and CLP+O3 (52,1 ± 8h), which did not show difference on survival compared to each other (p=0,4). CONCLUSION: In this rat model of sepsis, ozone therapy had a potential benefit in the modulation of inflammatory response and ALI, but no improvement on survival rates was observed.

Ao percorremos a trajetória de construção dos diagnósticos de enfermagem \"Trauma vascular\" e \"risco para trauma vascular\", buscamos, no capítulo 1, apresentar a classificação dos tipos de veias superficiais periféricas de adolescentes, adultos e idosos, segundo as características de uma veia passível de ser puncionada para fins terapêuticos e de diagnóstico, com base na aplicação da técnica Delphi, envolvendo juízes de quatro categorias profissionais distintas (angiologistas, anestesistas, enfermeiros e bioquímicos). Obtivemos índice de concordância para totalmente adequado/pertinente de mais de 90% e menos de 10% para moderadamente adequado/pertinente nos 13 critérios, a saber: mobilidade; trajeto; inserção/derivação; calibre; visibilidade; palpação e localização, tendo, como referencia: a articulação; localização da veia, tendo como referência, sua anatomia; regularidade do diâmetro do trajeto venoso; consistência do trajeto venoso; solução de continuidade das paredes do vaso; facilidade de punção e outros critérios a serem incluídos. No segundo capítulo, apresentamos a construção dos elementos (titulo, características definidoras, fatores relacionados) do diagnóstico de enfermagem \"trauma vascular\" e dos elementos (título e fatores de risco) para o diagnóstico de enfermagem \"Risco para trauma vascular\", baseando-nos na revisão literária e em nossa experiência profissional, e a validação de seus respectivos componentes, segundo o modelo de Fhering (1986) e adaptação do mesmo aos fatores relacionados e aos fatores de risco. Participaram 60 peritos. Reafirmamos: das 18 características definidoras analisadas, 15 são maiores (ponto de corte >=0,8) e 1 menor (ponto de corte >=0,50 ou <0,80); dos 14 fatores relacionados analisados, todos tiveram aceitação, sendo que 11 deles obtiveram escores de media ponderada >=0,8 e, dos 51 fatores de riscos analisados, 8 foram aprovados com escores >=0,50 para as situações ligadas a medicação e a forma/periodicidade de infusão; 4 foram aprovados para situações ligadas ao dispositivo endovenoso e seu tempo de permanência num mesmo sítio de inserção; 5 foram aprovados para situações ligadas a fixação do dispositivo endovenoso; 10 foram aprovados para situações ligadas ao indivíduo, seus hábitos, padrão de comunicação, estilo de vida e capacidade sensório-motora e 20 aprovados para as situações ligadas as decisões profissionais, a política institucional e ao procedimento propriamente. Finalmente, no capítulo 3, apresentamos validação clínica de alguns componentes do diagnóstico \"trauma vascular\" e \"Risco para trauma vascular\", utilizando um estudo de coorte, quando foram avaliadas 323 pessoas e 427 sítios de inserção de dispositivos endovenosos. Obtivemos significância no teste de ajuste do modelo para um conjunto de nove variáveis passíveis de serem transpostas para populações similares, merecendo destaque o tempo de permanência do dispositivo num mesmo sítio de inserção e a qualidade da fixação dos dispositivos (se fixos ou frouxos). Outros fatores mostraram-se relevantes somente para a população do estudo. / As we investigate the process of the construction of nursing diagnoses (vascular trauma and/or Risk of vascular trauma), we intend to classify, in the first chapter, the types of peripheral surface veins of teenage, adult, and elderly patients, according to the characteristics of a vein that can be punctured for therapeutic and diagnostics purposes, based on the application of the Delphi technique, judged by people from our different professional categories (angiologists, anesthetists, nurses and biochemists). We obtained a C.I. of over 90% for totally adequate/pertinent and less than 10% for moderately adequate/pertinent for the 13 criteria applied: mobility, course, insertion/derivation, caliber, visibility, palpation and localization with reference to the joint, location of the vein with reference to its anatomical structure, regulation of the diameter of the venous course, consistency of the venous course, continuity of the vessel tissue, how easy it is to puncture the vein, and other criteria to be included. In chapter 2, we present the construction of the elements (name, defining features, related factors) of the nursing diagnosis \"vascular trauma\" and of the elements (name, risk factor) of the nursing diagnosis \"risk of vascular trauma\", based on our reading of the existing literature and on our work experience, and the validation of their respective components as in Fhering´s (1986) model, and the adaptation of these to the related factors and risk factors. 60 experts took part. To summarize: of the 18 defining features analyzed, 15 were found to be greater (cut-off point >=0,8) and one smaller (cut-off point >=0,5 and <=0,8); of the 14 related factors analyzed, all met with approval, 11 of them with weighted average scores greater than or equal to 0,8 and of the 51 risk factors analyzed, 8 were approved with scores over 0,5 for situations connected with medication and the manner or intervals of infusion; 4 were approved for situations connected with the intravenous device and the time it remained in the same place of insertion, 5 were approved for situations connected with the individual, his/her habits, standard of communication, lifestyle and sensory-motor ability, and 20 were approved for situations connected with professional decisions, institutional policies and the procedure itself. Finally, in chapter 3, we present the clinical validation of some components of the diagnoses \"vascular trauma\" and \"risk of vascular trauma\", using a group study; our observations were based on 323 people and 427 point of insertion of the intravenous device. The results of the model adjustment test were significant tor a set of nine variables which may be transposed to similar populations. The most prominent of these variables were the time the device remained in one place of insertion and the quality of fixing of the devices (whether they were fixed or loose). Other factors were shown to be relevant only for the population under study.

Zielstellung: Ermittlung der Komplikationsrate nach intraartikulärer Punktion und Aufdeckung eines möglichen Zusammenhangs mit der Durchführung der Gelenkpunktion. Studiendesign: Es handelt sich um eine retrospektive Studie, basierend auf einer Fragebogenumfrage. Methoden: Erarbeitung eines Fragebogens und Versendung von 618 Exemplaren an 122 Pferdekliniken und 274 Fachtierärzte für Pferde (insgesamt 892 Fragebögen). Berücksichtigt wurden alle Pferdekliniken und Fachtierärzte für Pferde in Deutschland. Die Rückantwort erfolgte anonym. Insgesamt kamen 160 ausgefüllte Fragebögen zurück, von denen 155 in die statistische Auswertung einfließen konnten. Ergebnisse: Im Jahr 2006 wurden von 155 Tierärzten 65099 Gelenkpunktionen beim Pferd durchgeführt, das entsprach 420 Punktionen pro Tierarzt. Hierbei sind bei 51 Tierärzten insgesamt 93 Komplikationsfälle aufgetreten, was einer errechneten mittleren Komplikationsrate von 0,14 % entsprach. 64 (68,8 %) der Komplikationsfälle wurden geheilt, bei 13 (14,0 %) der Komplikationsfälle trat eine Besserung ein und sieben (7,5 %) mussten euthanasiert werden. Eine tödliche Komplikation trat somit zu 0,01 % (7 von 65099) nach einer Gelenkpunktion auf. Ein signifikanter Zusammenhang zwischen der mittleren Komplikationsrate und der Verwendung eines neuen Anbruches des zur Gelenkpunktion angewendeten Medikaments konnte festgestellt werden. Es konnte eine Tendenz zu einem Zusammenhang zwischen der mittleren Komplikationsrate und der Häufigkeit der Durchführung des Waschens vor der Punktion, zwischen der mittleren Komplikationsrate und des, zur Punktion verwendeten, Kanülendurchmessers sowie der mittleren Komplikationsrate und dem Ort der Punktion (Stall oder Klinik) festgestellt werden. Die meisten Punktionen wurden am Hufgelenk (25,0 %) und Fesselgelenk (24,4 %) durchgeführt. Hierauf folgten Tarsometatarsal- und Intertarsalgelenke (15,5 %), Kniegelenk (12,7 %), Talokruralgelenk (9,5 %), Karpalgelenk (7,7 %), Krongelenk (2,9 %), Schultergelenk (1,3 %), Ellbogengelenk (0,7 %) und Hüftgelenk (0,4 %). Die höchste mittlere Komplikationsrate hatte das Hufgelenk mit 0,28 %, dann folgten Ellbogengelenk (0,21 %), Karpalgelenk (0,16 %), Fesselgelenk (0,15 %), Talokruralgelenk (0,11 %), Kniegelenk (0,07 %), Krongelenk (0,05 %), und Tarsometatarsal- und Intertarsalgelenke (0,01 %). Beim Schulter- und Hüftgelenk traten keine Komplikationen auf. Beim Hufgelenk traten signifikant häufiger Komplikationen auf als bei den anderen Gelenken, außer dem Fesselgelenk. Beim Fesselgelenk traten signifikant häufiger Komplikationen auf als bei Tarsometatarsal- und Intertarsalgelenken. Schlussfolgerung und klinische Relevanz: Bei Gelenkpunktionen beim Pferd kann es mit geringer Wahrscheinlichkeit (0,14 %) zum Auftreten von Komplikationen kommen. Es wurde aufgezeigt inwiefern die, in der Literatur empfohlenen, Durchführungspunkte der Gelenkpunktion von den Praktikern umgesetzt wurden. Es wurde veranschaulicht, auf welche Schritte zur Verminderung des Komplikationsrisikos noch größerer Wert gelegt werden sollte. Die Komplikationsanfälligkeit ist zwischen den Gelenken verschieden, wobei vor allem das Hufgelenk mit einem größeren Risiko belastet zu sein schien, was hier ein besonders sorgfältiges Vorgehen verlangt. Die Studie lieferte erstmals Aussagen zum Komplikationsauftreten nach Gelenkpunktion beim Pferd, auch bezüglich der einzelnen Gelenke. Die Ergebnisse können als Grundlage zur Besitzerinformation dienen und hilfreich für die Gutachtertätigkeit sein.

<p>Sepsis and septic shock are life-threating conditions, which resulted from a continuum of the body’s response to overwhelming infection. Elimination of bacteria through antibiotics is not sufficient, because the host is still left with a large amount of lipopolysaccharide (LPS) that prevents the host immune system from returning to normal homeostasis. Synthetic LPS antagonists that can bind to LPS via electrostatic and/or hydrophobic interactions cause systemic toxicities. Moreover, LPS elimination alone may not address already established complications of sepsis. To address these challenges, we propose to develop nanoparticle formulations of LPS antagonists (D-TZP) that can be delivered systemically. Specifically, cholecalciferol (vitamin D) was encapsulated in a self-assembly of tannic acid/Fe³⁺ coordination complex (pTA) capsule, forming a core that could be surface-modified with LPS adsorbents, such as low molecular weight succinylated chitosan (LMZWC) and polymyxin B (PMB). D-TZP suppressed pro-inflammatory effects of LPS on the engineered human monocytes with significantly less cytotoxicity than free PMB at the equivalent dose. D-TZP increased the maximum tolerated dose of PMB by both intraperitoneal and intravenous administration. In the LPS-induced mouse model of sepsis, systemic administration of D-TZP immediately after LPS challenge neutralized the lethal effect of LPS. D-TZP also reduced the mortality of mice when given 2 h after the LPS challenge. D-TZP inhibited the mortality in the cecal ligation and puncture (CLP)-induced bacteremia mouse model when given IV 2 h after the insult. In the CLP model, the D-TZP-treated animals also showed lower levels of both TNF-α and IL-10 cytokines as well as D-dimer levels, reflecting the attenuation of disseminated intravascular coagulation, compared to the vehicle-treated control group. Collectively, these results support that the D-TZP is a safe and effective systemic intervention of sepsis.<br></p>

Sepsis is a clinical syndrome characterized by the systemic activation of inflammatory and coagulation pathways in response to microbial infection of normally sterile parts of the body. Despite considerable advances in our understanding of sepsis pathophysiology, sepsis remains the leading cause of death in non-coronary intensive care units (ICU) with a global disease burden between 15 and 19 million cases per year (Dellinger et al., 2008). Severe sepsis, defined as sepsis associated with organ dysfunction is associated with mortality rates of 33% to 45%. The incidence of severe sepsis continues to increase by 1.5% per annum due to the aging population, a rise in the prevalence of comorbidities, and the wider use of immunosuppressive agents and invasive procedures (Angus et al., 2001). Over the past several decades, many potential treatments for sepsis have shown early promise, yet have failed to improve survival in over 100 Phase II and Phase III clinical trials (Marshall, 2014) suggesting that some fundamental knowledge is lacking in our understanding of sepsis pathophysiology. Emerging studies on cell-free DNA (cfDNA), DNA released extracellularly into the circulation, demonstrate that cfDNA is a crucial link between inflammation and coagulation . In various conditions characterized by excessive inflammatory responses or aberrant prothrombotic responses, cfDNA has been implicated in exacerbating disease pathology (Atamaniuk, Kopecky, Skoupy, Säemann, & Weichhart, 2012; Fuchs, Brill, & Wagner, 2012; Swystun, Mukherjee, & Liaw, 2011). In clinical sepsis, levels of cfDNA upon admission into the ICU have strong prognostic value in predicting mortality (Dwivedi et al., 2012; Saukkonen et al., 2008). However, it is unclear whether these increases in cfDNA are an epiphenomenon during sepsis progression, or whether cfDNA actively plays a role in sepsis pathophysiology. In this work, in vivo studies were conducted to characterize the role of cfDNA in sepsis, the effects of DNase administration, and the potential mechanism by which cfDNA is released during experimental sepsis. In addition, mortality studies were conducted to identify surrogate markers of death to promote the design of humane and ethical animal studies in conducting sepsis research. Polymicrobial sepsis was induced via a surgical procedure whereby the cecum is exteriorized, ligated and punctured twice to introduce a continuous source of microorganisms, a model termed cecal ligation and puncture (CLP). In our CLP sepsis model, levels of cfDNA increased in a time-dependent manner. These increases accompanied an early pro-inflammatory response marked by increased pro-inflammatory IL-6, a transient increase in anti-inflammatory IL-10, and elevated lung myeloperoxidase (MPO) activity. Septic mice with elevated cfDNA levels also had high bacterial loads in the lungs, blood, and peritoneal cavity fluid. Organ damage was also observed in mice following CLP surgery versus mice subjected to the non-septic sham control surgery marked by increased levels of creatinine and alanine aminotransferase (ALT) indicative of kidney and liver injury, respectively. Histological analyses further confirmed lung and kidney damage following CLP surgery. Changes in coagulation were also observed in septic mice as mice subjected to CLP had sustained increases in thrombin-antithrombin (TAT) complexes. In addition, plasma from CLP-operated mice had increased thrombin generation (i.e. increased endogenous thromin potential, increased peak thrombin, decreased time to peak, and decreased lag time) mediated by FXIIa and enhanced by platelets. Following CLP-induced sepsis, elevations in cfDNA levels accompanied pro-inflammatory and pro-coagulant responses. The effects of in vivo DNase treatment in septic mice were time-dependent. Early DNase treatment when cfDNA levels were low resulted in an exaggerated pro-inflammatory response marked by increased plasma IL-6 levels and increased lung damage. In contrast, delayed DNase treatment at time-points when cfDNA levels were elevated suppressed inflammation characterized by an increase in anti-inflammatory IL-10 and reductions in cfDNA, IL-6, lung MPO, and ALT activity. Furthermore, delayed DNase administration resulted in decreased bacterial load in the lungs, blood, and peritoneal cavity fluid. Delayed DNase treatment also resulted in blunted pro-coagulant responses as levels of TAT complexes were suppressed and thrombin generation from septic mouse plasma was normalized. Moreover, DNase treatment when cfDNA levels were elevated increased survival in CLP-operated mice by 80% and reduced lung and liver damage. These findings suggest that administration of DNase when cfDNA levels are elevated may reduce pro-inflammatory and pro-coagulant responses and that delayed DNase treatment may infer protection in the CLP model of sepsis. One mechanism by which cfDNA is released is via the formation of neutrophil extracellular traps (NETs). Upon inflammatory stimulation, some neutrophils release chromatin material and antimicrobial proteins (i.e. neutrophil elastase, MPO, and histones) in an active process termed NETosis. Although NETs ensnare bacteria and exert antimicrobial properties, NETs may also exert harmful effects on the host by activating inflammation and coagulation. While some in vitro evidence suggest that neutrophils are the main source of cfDNA released following inflammatory stimulation, others have reported that neutrophils are not the main source of circulating cfDNA following septic challenge. To determine whether NETs contribute to cfDNA released during CLP sepsis, genetically modified mice that are incapable of forming NETs, PAD4-/- mice, were used. Levels of cfDNA in PAD-/- mice were significantly lower than cfDNA levels in C57Bl/6 mice following CLP surgery, suggesting that NETs were a source of cfDNA in our model. Levels of IL-6, MPO, and bacterial load in the lungs, blood, and peritoneal cavity were significantly reduced, indicating that NETs exert pro-inflammatory effects in CLP sepsis. Thrombin generation was also suppressed in PAD4-/- mice which suggests that NETs contribute to thrombin generation following CLP sepsis. NETs contribute to increases in circulating cfDNA and may exacerbate pathology by driving pro-inflammatory and pro-coagulant responses in CLP-induced sepsis. Appreciating the implications of conducting research using animals, it is pertinent that researchers ensure the highest ethical standards and design animal studies in the most humane, yet scientifically rigorous manner. Using mortality studies, we validated the utility of physiological and phenotypic markers to assess disease severity and predict death in murine sepsis. Temperature via a rectal probe monitor and sepsis scoring systems which assess components such as orbital tightening, level of consciousness, and activity were effective surrogate markers of death. These tools offer a non-invasive assessment of disease progression which do not artificially exacerbate sepsis pathology and immediate information regarding any changes in the health status. Surrogate markers of death also provide reliable monitoring to meet increasing standards of ethical, humane animal research and a feasible and cost-efficient means to obtain vital signs in small rodents. We have proposed a scoring system which can be used for assessing disease severity, endpoint monitoring, and predicting death to obviate inhumane methods of using death as an endpoint in sepsis studies. In summary, cfDNA levels are elevated in CLP-induced sepsis and these elevations accompany pro-inflammatory and pro-coagulant responses. NETosis may be a mechanism by which cfDNA is released and NETs may drive inflammation and coagulation in CLP sepsis. Delayed DNase administration may suppress inflammation and coagulation and may be protective in polymicrobial sepsis. In future animal sepsis studies, surrogate markers of death and a sepsis scoring system can be used in place of death as an endpoint to raise the standards in conducting ethical, humane sepsis research. / Thesis / Doctor of Philosophy (PhD)

Objectives set by the EU means that all buildings after 2020 has to be nearly zero energy buildings. This means that thicker layers of insulation have to be added in the wall construction which makes the wall thicker. It means that the living area will be reduced. Vacuum insulation is a highly effective type of insulation and because of its low thermal conductivity it has the ability to reduce the thickness in wall structures. This project investigates a proposal to apply vacuum insulation in one-storey buildings. In order to achieve the goals of the project, a proposal for a one-storey building was developed. Calculations have been made and the proposal was developed as an alternative to show how to construct a family home containing vacuum insulation. The empirical data was collected through interviews, document analysis and literature studies. The collected data was analyzed together with the theoretical framework that has been developed through literature studies and document analysis. Creating a wall construction containing vacuum insulation as a primary insulation usually means that the wall will be considerably thinner than a wall construction with traditional insulation. This means that living area can be saved. Vacuum insulation has to be protected properly as it is easily punctured where upon it loses the most of its insulation capacity. Vacuum insulation is not common on the Swedish construction market today, this is due to many factors, including its high price. Vacuum insulation is a good problem solver which can be used in bay windows to gain extra space. One can also make use for it in tight spaces. From an economic point of view vacuum insulation offers the greatest advantages in cities where living space is considerably higher than in rural areas. To take part of the work there is no need for prior knowledge about vacuum insulation. The project focuses only on wall structures in the single-storey villas, therefor, no indentations has been made on the floor- and roof structures or other building types. The project only focuses on newly constructed buildings. No calculations are made for moisture or production costs. / Mål uppsatta av EU innebär att samtliga byggnader som uppförs vid år 2020 måste vara nära-nollenergihus. För väggarna i konstruktionen innebär det att tjockare lager av isolering måste adderas vilket ger bredare väggkonstruktioner. Bredare väggkonstruktioner innebär även att boarean minskas. Vakuumisolering är ett högeffektivt isoleringsmaterial som genom sin låga värmeledningsförmåga har möjligheten att minska tjockleken vid väggkonstruktioner på grund av dess tunna skikt. Arbetet utreder ett förslag att applicera vakuumisolering i enplansvillor. För att uppnå arbetets mål har ett förslag på enplansvilla tagits fram. Beräkningar har gjorts och förslaget är framtaget som ett alternativ för att visa hur en villa innehållande vakuumisolering kan utformas. Det empiriska materialet har samlats in genom intervjuer, dokumentanalyser samt litteraturstudier. Empirin analyseras sedan tillsammans med det framtagna teoretiska ramverket genom litteraturstudier och dokumentanalyser. Att skapa en väggkonstruktion med vakuumisolering som primär isolering betyder oftast att väggen blir avsevärt mycket tunnare än en väggkonstruktion av traditionell isolering, vilket betyder att boarea kan sparas. Vakuumisolering måste skyddas på rätt sätt i väggkonstruktioner eftersom materialet lätt punkteras varpå det förlorar den största delen av sin isoleringsförmåga. Idag är inte vakuumisolering utbrett på den svenska byggmarknaden vilket beror på många faktorer, bland annat dess höga pris. Vakuumisolering är en väldigt bra problemlösare som med fördel kan användas i burspråk för att vinna extra utrymme. Det kan även användas i trånga utrymmen som elnischar. Ur ekonomisk synpunkt ger vakuumisolering störst fördel i städer där boarea per kvadratmeter är högre än motsvarande på landsbygden. För att ta del av arbetet krävs inga förkunskaper om vakuumisolering. Arbetet fokuserar endast på väggkonstruktioner i enplansvillor, därför har inga fördjupningar skett på golv- och takkonstruktioner eller andra byggnadstyper. Enbart nybyggnationer av trästommar är utrett. Beräkningar är inte gjorda för fukt och produktionskostnader.

En chirurgie vasculaire, l’accès à l’artère fémorale, qu’il soit par une incision chirurgicale ou par une approche percutanée, est très fréquemment utilisé pour une multitude d’interventions vasculaires ou endovasculaires; pour des pontages divers, le traitement d’occlusions artérielles, la réparation d’anévrismes et la pose d’endoprothèses. L’objectif général de ce projet de recherche est de faciliter et réduire les risques des approches de l’artère fémorale par une meilleure compréhension anatomique du triangle fémoral. La méthodologie a été réalisée grâce à l’utilisation de cadavres spécialement embaumés par la méthode développée par Walter Thiel. Les résultats présentés dans ce mémoire ont permis de proposer des solutions en réponse à des problèmes cliniques en chirurgie vasculaire. Dans un premier temps, l’étude de la vascularisation cutanée du triangle fémoral a mené à proposer de nouvelles incisions chirurgicales afin de limiter la dévascularisation cutanée des plaies et ainsi réduire les problèmes de cicatrisation observés. Ensuite, nous avons validé l’identification radiographique et échographique de l’artère fémorale à son croisement avec le ligament inguinal afin de faciliter l’identification d’un site de ponction artérielle adéquat. Enfin, nous avons développé une méthode échographique simple qui facilite l’approche percutanée de l’artère fémorale, même chez les patients obèses. Les retombées de ce projet de recherche sont multiples pour les cliniciens, l’étude fournit une meilleure compréhension anatomique tridimensionnelle du triangle fémoral et les techniques proposées dans ce mémoire pourront apporter une amélioration de la pratique chirurgicale et faciliter le travail des médecins. Toutefois, ces propositions devront maintenant être validées en clinique. / In vascular surgery, access to the femoral artery is frequently used either through a surgical incision of the groin or by a percutaneous approach in a wide variety of vascular and endovascular procedures; for multiple bypasses, treatment of arterial occlusions, aneurysms repair and placement of various stents. The general purpose of this study is to facilitate and reduces the risks of both the surgical and the percutaneous approaches of the femoral artery through a better anatomical understanding of the femoral triangle. The methodology was conducted on specifically embalmed cadavers according to the method developed by Walter Thiel. The results presented in this memory allowed us to propose solutions to clinical problems in vascular surgery. First, the study of the cutaneous vascularisation of the femoral triangle led to suggest new surgical approaches in order to reduce lesions of the cutaneous arteries and the potential devascularization of the borders of the wound yielding a better postoperative outcome. Then, we validated the radiographic and ultrasonographic position and identification of the inguinal ligament and the proximal femoral artery to facilitate the identification of a safe arterial puncture site. Moreover, we developed a simple method for ultrasound-guided arterial puncture, suitable for obese patients, to facilitate the percutaneous approach of the femoral artery. The benefits of this research project are multiple for clinicians, the study provides a better tridimensional anatomic understanding of the femoral triangle and the techniques proposed will lead to an improved surgical practice and facilitate the work of doctors. However, our work and propositions still need to be validated clinically.

O recrutamento de neutrófilos para o sítio da infecção é um evento crucial para o combate aos microrganismos e sobrevivência na sepse. A migração destes polimorfonucleares é dirigida através de um gradiente quimiotático por meio do reconhecimento de quimiocinas por receptores acoplados a proteína G (GPCRs), os quais são regulados por quinases específicas (GRKs). Estudos prévios demonstraram que na sepse ocorre uma falência na migração de neutrófilos para o foco infeccioso em função da dessensibilização de receptores quimiotáticos via GRKs induzida pela ativação de receptores toll-like (TLRs), TLR2 e TLR4. Apesar de a ausência de TLR9 em células dendriticas ter sido relacionada a maior sobrevivência de camundongos sépticos, o papel do TLR9 atuando diretamente em neutrófilos não foi avaliado. Objetivando preencher esta lacuna, propôs-se avaliar o papel direto de TLR9 na falência de migração de neutrófilos na sepse. Os camundongos TLR9-/- apresentaram maior sobrevivência a sepse polimicrobiana avaliada por meio do modelo de ligadura e perfuração do ceco (CLP). A deficiência de TLR9 também acarretou em aumento na migração de neutrófilos para o foco da infecção, menor seqüestro de neutrófilos no pulmão, bem como, menor número de bactérias no lavado peritoneal e sangue. A ativação de TLR9 por oligodeoxinucleotídeo contendo o dinucleotídeo CpG não metilado (ODN CpG) nos neutrófilos reduziu a quimiotaxia destes em direção a quimiocina CXCL2 e expressão do receptor quimiotático CXCR2. Além disso, neutrófilos estimulados com ODN CpG apresentaram aumento na expressão da quinase tipo 2 relacionada a receptores acoplados a proteína G (GRK2). Dessa forma, a ativação de TLR9 em neutrófilos circulantes no sangue é prejudicial na sepse por reduzir a quimiotaxia destes para o foco da infecção ao induzir a dessensibilização de CXCR2 via GRK2. / The recruitment of neutrophils to the site of infection is a crucial event for combating the microorganisms and survival on sepsis. The neutrophil migration is directed by a chemotactic gradient through the recognition of chemokines by G protein-coupled receptors (GPCRs), which are regulated by specific kinases (GRKs). Previous studies have shown a failure of neutrophil migration into infectious focus on sepsis due to chemotactic receptor desensitization via GRKs induced by activation of toll- like receptors (TLRs), TLR2 and TLR4. Despite the absence of activation of TLR9 in dendritic cells have been related to increase survival of septic mice, the role of TLR9 acting directly on neutrophils was not evaluated. We proposed to verify the direct role of TLR9 in the failure of neutrophil migration on sepsis. The TLR9 knockout mice (TLR9-/-) showed high survival to polymicrobial sepsis using cecal ligation and puncture model (CLP). TLR9-/- mice had high neutrophil migration to the focus of infection, low neutrophil sequestration in the lung, as well as, few bacteria in the peritoneal exudates and blood. The activation of TLR9 by oligodeoxinucleotide containing unmethylated dinucleotide CpG (CpG ODN) in neutrophils also reduced chemotaxis toward CXCL2 and the expression of chemokine receptor CXCR2. In addition, neutrophils stimulated with CpG ODN showed increased expression of kinase-related G protein-coupled receptor type 2 (GRK2). Thus, the activation of TLR9 in blood circulating neutrophils is harmful on sepsis by reducing their chemotaxis into the site of the infection by inducing CXCR2 desensitization via GRK2.

O recrutamento de neutrófilos para o sítio da infecção é um evento crucial para o combate aos microrganismos e sobrevivência na sepse. A migração destes polimorfonucleares é dirigida através de um gradiente quimiotático por meio do reconhecimento de quimiocinas por receptores acoplados a proteína G (GPCRs), os quais são regulados por quinases específicas (GRKs). Estudos prévios demonstraram que na sepse ocorre uma falência na migração de neutrófilos para o foco infeccioso em função da dessensibilização de receptores quimiotáticos via GRKs induzida pela ativação de receptores toll-like (TLRs), TLR2 e TLR4. Apesar de a ausência de TLR9 em células dendriticas ter sido relacionada a maior sobrevivência de camundongos sépticos, o papel do TLR9 atuando diretamente em neutrófilos não foi avaliado. Objetivando preencher esta lacuna, propôs-se avaliar o papel direto de TLR9 na falência de migração de neutrófilos na sepse. Os camundongos TLR9-/- apresentaram maior sobrevivência a sepse polimicrobiana avaliada por meio do modelo de ligadura e perfuração do ceco (CLP). A deficiência de TLR9 também acarretou em aumento na migração de neutrófilos para o foco da infecção, menor seqüestro de neutrófilos no pulmão, bem como, menor número de bactérias no lavado peritoneal e sangue. A ativação de TLR9 por oligodeoxinucleotídeo contendo o dinucleotídeo CpG não metilado (ODN CpG) nos neutrófilos reduziu a quimiotaxia destes em direção a quimiocina CXCL2 e expressão do receptor quimiotático CXCR2. Além disso, neutrófilos estimulados com ODN CpG apresentaram aumento na expressão da quinase tipo 2 relacionada a receptores acoplados a proteína G (GRK2). Dessa forma, a ativação de TLR9 em neutrófilos circulantes no sangue é prejudicial na sepse por reduzir a quimiotaxia destes para o foco da infecção ao induzir a dessensibilização de CXCR2 via GRK2. / The recruitment of neutrophils to the site of infection is a crucial event for combating the microorganisms and survival on sepsis. The neutrophil migration is directed by a chemotactic gradient through the recognition of chemokines by G protein-coupled receptors (GPCRs), which are regulated by specific kinases (GRKs). Previous studies have shown a failure of neutrophil migration into infectious focus on sepsis due to chemotactic receptor desensitization via GRKs induced by activation of toll- like receptors (TLRs), TLR2 and TLR4. Despite the absence of activation of TLR9 in dendritic cells have been related to increase survival of septic mice, the role of TLR9 acting directly on neutrophils was not evaluated. We proposed to verify the direct role of TLR9 in the failure of neutrophil migration on sepsis. The TLR9 knockout mice (TLR9-/-) showed high survival to polymicrobial sepsis using cecal ligation and puncture model (CLP). TLR9-/- mice had high neutrophil migration to the focus of infection, low neutrophil sequestration in the lung, as well as, few bacteria in the peritoneal exudates and blood. The activation of TLR9 by oligodeoxinucleotide containing unmethylated dinucleotide CpG (CpG ODN) in neutrophils also reduced chemotaxis toward CXCL2 and the expression of chemokine receptor CXCR2. In addition, neutrophils stimulated with CpG ODN showed increased expression of kinase-related G protein-coupled receptor type 2 (GRK2). Thus, the activation of TLR9 in blood circulating neutrophils is harmful on sepsis by reducing their chemotaxis into the site of the infection by inducing CXCR2 desensitization via GRK2.

L'encéphalopathie associée au sepsis (EAS) est définie comme un dysfonctionnement cérébral diffus induit par une réponse systémique à une infection. Chez les patients septiques, l'imagerie par résonance magnétique (IRM) a indiqué à la fois des anomalies de la substance grise (SG) et blanche (SB) associées à des troubles cognitifs graves, y compris le delirium. Pour améliorer notre compréhension des changements hémodynamiques, métaboliques et structuraux associés au sepsis, différentes séquences d'IRM ont été réalisées chez des rats ayant subi une injection ip de solution saline ou de lipopolysaccharide bactérien (LPS) 2,5h plus tôt ou une ligature et ponction caecale 24h plus tôt. Après ip LPS, l'IRM de contraste de phase a été réalisée pour étudier le flux des artères cérébrales antérieures et moyennes et le marquage des spins artériels (ASL) pour étudier la perfusion des structures cérébrales de la SB et SG. Des séquences d'imagerie par diffusion pondérée (DWI) ont été utilisées pour évaluer les changements structurels. Après la chirurgie CLP, ASL a été utilisé pour étudier les changements de la microcirculation. L'imagerie pondérée en T2, l'imagerie du tenseur de diffusion (DTI) et les statistiques spatiales basées sur les faisceaux (TBSS) ont été réalisées pour caractériser les événements structurels dans différentes structures cérébrales. Après imagerie, les animaux ont été sacrifiés et leur cerveau a été traité pour l'histologie afin de détecter l'enzyme synthétisant les prostaglandines vasoactives cyclooxygénase-2 (COX-2) et le canal hydrique astrocytaire aquaporin-4 (AQP4) dont l'expression peut être régulée à la hausse, évaluer la présence d'immunoglobulines périvasculaires (Ig) indiquant une rupture de la barrière hémato-encéphalique (BHE) et étudier la morphologie des glies puisque la microglie et l’astroglie changent de morphologie lors des conditions inflammatoires. L'IRM n'a indiqué aucun changement hémodynamique dans la substance grise après l'administration de ip LPS, alors qu'une perfusion cérébrale accrue a été montrée au niveau du corps calleux comme indiqué par l'ASL. DTI a indiqué une augmentation de la diffusion des molécules d’eau parallèlement aux fibres du corps calleux. Ces changements étaient accompagnés d'une dégradation de BHE dans la SB ainsi que la substance grise corticale et striatale adjacente tel est indiqué par la présence périvasculaire d'IgG, sans aucun changement majeur de COX-2 vasculaire ou de morphologie des glies du coprs calleux. Le dysfonctionnement du SNC induit par le sepsis a résulté en une augmentation du contraste pondéré en T2 dans le cortex, le striatum et la base du cerveau, une diminution de la perfusion sanguine dans le cortex et une augmentation de la diffusion hydrique du corps calleux et du striatum ventral. Ces changements ont été associés dans la SB à des modifications de la morphologie des glies et dans la substance grise à une expression constitutive de COX-2 et AQP4 plus faible dans le cortex cérébral. La comparaison entre CLP ayant subit ou non une IRM sous anesthésie à l'isoflurane a montré une réponse inflammatoire réduite tel est indiqué par l'expression de COX- 2, une activation réduite des glies ainsi qu’une lésion réduite de la BHE dans le CLP subissant une IRM sous anesthésie. Collectivement, nos résultats suggèrent que des changements hémodynamiques peuvent survenir en l'absence de flux altéré dans les artères irriguant le cerveau antérieur. Ensuite, l'altération de la structure de la SB est une étape précoce de la pathogenèse de l’EAS qui peut résulter soit de la dégradation de la BHE, soit de l'activation des glies. Cette étude sous-tend l'effet délétère d'une seule exposition à l'anesthésie à l'isoflurane qui peut être atténuée par une seconde exposition chez les rats ayant subi une laparotomie ainsi que les effets de l'inflammation systémique induite par le CLP sur les glies pouvant être atténués par imagerie sous anesthésie à l'isoflurane. / Sepsis-associated encephalopathy (SAE) refers to central nervous system dysfunction during the systemic inflammatory response to infection. In septic patients with encephalopathy MRI has indicated both gray and white matter abnormalities that were associated with worse cognitive outcome including delirium. To improve our understanding of sepsis-associated hemodynamic, metabolic, and structural changes, different MRI sequences were performed in rats that either underwent an i.p injection of saline or bacterial lipopolysaccharide (LPS) 2.5h earlier or cecal ligation and puncture (CLP) 24h earlier. After ip LPS, phase contrast MRI was performed to study anterior and middle cerebral arteries flow and Arterial Spin Labeling (ASL) to study perfusion of white and grey matter brain structures. Diffusion Weighted Imaging (DWI) sequences was used to assess structural changes. After CLP surgery, ASL was used to study microcirculation changes. T2-Weighted Imaging, Diffusion Tensor Imaging (DTI) and tract-based spatial statistics (TBSS) were performed to characterize structural events in different brain structures. After imaging, animals were sacrificed and their brains processed for histology to detect the vasoactive prostaglandin-synthesizing enzyme cyclooxygenase-2 (COX-2) and the astrocytic aquaporin-4 water channel (AQP4) the expression of which can be upregulated during inflammation, to assess the presence of perivascular immunoglobulins (Ig) indicating blood-brain barrier (BBB) leakage and to study glia cell morphology as both microglia and astrocytes are known to change their morphology in inflammatory conditions. Magnetic resonance rat brain imaging indicated no hemodynamic changes in the grey matter after ip LPS administration while an increased CBF was shown in corpus callosum white matter as indicated by ASL. DTI indicated increased water diffusion parallel to fibers of the corpus callosum white matter. These changes were accompanied by BBB breakdown in the white matter and adjacent cortical and striatal grey matter as indicated by the perivascular presence of IgG, but no major changes in vascular COX-2 or white matter glia cell morphology. CLP induced sepsis-associated CNS dysfunction resulted in higher T2-weighted contrast intensities in the cortex, striatum and base of the brain, decreased blood perfusion distribution to the cortex and increased water diffusion in the corpus callosum and ventral striatum compared to sham surgery. These changes were associated in the white matter with modifications in glia cells morphology and in the grey matter with lower expression of constitutive COX-2 expression and AQP4 in the cerebral cortex. The comparison between CLP that underwent or not MRI under isoflurane anesthesia indicated reduced inflammatory response as indicated by COX-2 expression, reduced glia activation and reduced BBB damage in CLP that underwent MRI under isoflurane anesthesia. Collectively, our results suggest that hemodynamic changes may occur in the absence of altered flow in forebrain irrigating arteries. Then, altered white matter structure is an early step in SAE pathogenesis that may result either from BBB breakdown or glial cells activation. This study underlies the deleterious effects of a single exposure to isoflurane anesthesia that may be mitigated by a second exposure in sham-operated rats and the effects of CLP-induced systemic inflammation on glial cells that can be attenuated by imaging under isoflurane anesthesia.