Les maladies thrombotiques sont la cause majeure de décès dans les pays industrialisés. Les cellules souches dérivées de la moelle osseuse ont été impliqués dans la réparation vasculaire et contribuent à restaurer l'intégrité de l'endothélium. Les cellules progéniteurs CD34 positives ont été rapportées pour jouer un rôle important suite à une blessure de la paroi vasculaire en se liant aux plaquettes ou à la fibrine, modulant la formation du thrombus et participant à la ré-endothélialisation de la paroi vasculaire lésée. Parmi les cellules progéniteurs CD34 positives, les cellules endothéliales formant colonie (ECFC) ont été caractérisés comme présentant des propriétés endothéliale. La première partie de ce travail met en évidence un nouveau partenariat entre les neutrophiles et les ECFC. Par l'utilisation de la microscopie intravitale chez les souris, nous montrons que les neutrophiles recrutent les ECFC au site de lésion vasculaire induite par rayon laser. Ce recrutement est dépendant du PSGL-1 mais indépendant de l'expression de la P-sélectine. L'interaction avec les neutrophiles accroît le potentiel pro-angiogénique des ECFC in vitro. L'identification de ce nouveau partenariat entre les neutrophiles et les ECFC dans la formation de thrombus possède des implications potentiellement critiques dans le contrôle de l'angiogenèse.
La seconde partie de ce travail décrit une nouvelle méthode pour imager en temps réel le devenir des ECFCs in vivo. Nos résultats montrent que les ECFC s'accumule au site de lésion vasculaire.
En conclusion, ce travail démontre que les ECFC peuvent participer favoriser et réguler l'angiogenèse en interagissant avec les neutrophiles. / Thrombotic diseases are major cause of death in industrialized countries. Bone marrow derived progenitor cells have been implicated in vascular repair and contribute to restore the integrity of endothelium, thus constituting important partners for vascular wall restoration. CD34 positive progenitors cells were reported to play an important role following an injury of the vessel wall by binding to platelets or fibrin, modulating thrombus formation and participating in the re-endothelization of the injured vessel wall. Among CD34 positive progenitors cells, Endothelial Colony forming Cells (ECFCs) have been characterized as a unique subset displaying endothelial properties. The first part of this work described a new partnership between neutrophils and ECFCs. Using high- speed digital fluorescent intravital microscopy in living mice we show that neutrophils recruit ECFCs at the site of a laser-induced vessel injury via PSGL-1 axis independently of P-selectin. This interaction enhances the pro-angiogenic potential of ECFCs in vitro. The identification a new central partnership between neutrophils and ECFC in thrombus formation has critical potential implications to control angiogenesis.
The second part of this work, described a new method to image in real-time the homing and survival ECFCs in vivo. Our results show that ECFCs transducted with a gene coding for luciferase accumulates at site of vascular traumatism.
In conclusion, this work indicates that ECFCs may participate in the promotion and regulation of angiogenesis by interacting with neutrophils at a site of vascular traumatism.

L’hémostase est un processus physiologique permettant de préserver l’intégrité du système vasculaire et de prévenir une perte de sang en réponse à une blessure. En situation pathologique, comme dans le cas de cancers, d’infections ou de maladies cardiovasculaires, il peut y avoir activation de la cascade de coagulation entraînant la formation d’un thrombus.Dans cette étude, nous avons utilisé la microscopie intravitale dans un modèle de blessure au rayon laser pour comprendre les mécanismes cellulaires et moléculaires mis en jeu dans la formation d’un thrombus plaquettaire en conditions physiologiques et pathologiques lors du développement d’un cancer. La première partie de ce travail a consisté à décrire l’implication des polynucléaires neutrophiles dans la formation d’un thrombus. Nous montrons que les neutrophiles sont les premières cellules à s’accumuler au niveau du site de blessure et représentent la principale source de facteur tissulaire menant à la génération de fibrine et à la formation du thrombus. Ces neutrophiles sont nécessaires au recrutement de cellules endothéliales progénitrices (Endothelial colony- forming cells, ECFCs), qui sont des cellules capables de jouer un rôle important dans la réparation vasculaire. Dans une seconde partie, nous avons déterminé, dans des modèles murins adaptés, l’implication de l’activation du FT et des plaquettes dans la thrombose associée au cancer. En conclusion, notre travail donne de nouvelles perspectives dans la compréhension du rôle pathophysiologique des neutrophiles, des cellules progénitrices endothéliales et des plaquettes. / Hemostasis is a physiological process to preserve the integrity of the vascular system and to prevent blood loss in response to injury. In pathological conditions, such as cancers, infections or cardiovascular diseases, the blood coagulation cascade can be activated, leading to the formation of a platelet thrombus.Using a laser-injury model coupled with a high-definition, high-speed camera, we explored the cellular and molecular mechanisms involved in thrombus formation in physiological and in pathological conditions associated with the development of a cancer. The first part of this work describes the role of polymorphonuclear neutrophils (PMNs) in thrombus formation. We show that PMNs are the first cells to accumulate at the site of injury and represent the main source of blood-borne tissue factor (TF), leading to the generation of fibrin and thrombus formation. We also show that once present at the site of injury, PMNs recruit Endothelial Progenitor Cells (endothelial colony-forming cells, ECFCs), which play a key role in vascular repair. The second part of this work we determined, in dedicated mouse models, the involvement of TF and platelet activation in thrombosis associated with cancer. Together, our findings provide new perspectives in the understanding of the pathophysiological role of polymorphonuclear neutrophils, Endothelial Progenitor Cells and platelets.

Os progestagênios isolados são contraceptivos indicados que em trombofílicas e/ou com passado de tromboembolismo venoso (TEV). Algumas dessas mulheres, também utilizam anticoagulantes orais (ATCO) o que pode implicar em alterações no padrão de sangramento menstrual associado ao uso de progestagênios isolados. Objetivo: avaliar os efeitos do uso de ATCO no padrão de sangramento associado ao sistema intrauterino liberador de levonorgestrel (SIU-LNG) em mulheres trombofílicas e/ou com passado de TEV . Métodos: Trata-se de um estudo de coorte prospectiva que dividiu as mulheres trombofílicas e/ou com passado de TEV em dois grupos, usuárias de ATCO e não-usuárias de ATCO. Padrão de sangramento, peso, índice de massa corpórea (IMC) e circunferência abdominal foram comparados entre os grupos antes, 6 e 12 meses após a inserção do SIU-LNG. Resultados: Foram incluídas 33 mulheres, entre 18 e 45 anos, 16 usuárias de ATCO e 17 não-usuárias. Houve aumento de 3,9% do peso e 3,8% do IMC em das usuárias de ATCO após 12 meses da inserção do SIU-LNG (p<0,01). Houve aumento de hemoglobina e hematócrito em ambos os grupos. Não se observou diferença entre o padrão de sangramento de ambos os grupos, sendo a amenorréia o padrão mais frequente nos dois grupos (41,2% em ambos) 12 meses após a inserção do SIU-LNG. Utilizar ATCO não aumentou a freqüência e a duranção de sangramento genital. Conclusão: As usuárias e não-usuárias de ATCO tiveram padrão de sangramento semelhante após a inserção do SIU-LNG. Os níveis de hemoglobina e hematócrito aumentaram em ambos os grupos / Background: Progestogen-only contraceptives (POCs) are suitable for women with thrombophilia and/or a history of venous thromboembolism (VTE). Several of these women, however, use oral anticoagulant therapy (OAT), which can impair the bleeding pattern associated with POC use. We evaluated the effects of OAT use on the bleeding pattern associated with the levonorgestrel-releasing intrauterine system (LNG-IUS) in women with thrombophilia and/or a history of VTE. Study Design: This prospective cohort study followed 2 groups of women, all of whom were thrombophilic and/or had a history of VTE: OAT users and non-users. Bleeding patterns, blood pressure, body mass index (BMI), weight, complete blood count and waist circumference were compared between the 2 groups before and 6 and 12 months after LNG-IUS insertion. Results: The patient cohort consisted of 33 women aged 18 to 45 years old, including 16 OAT users and 17 non-users. Body weight increased by 3.9% and BMI by 3.8% in OAT users 12 months after LNG-IUS insertion (p< 0,01). Hemoglobin and hematocrit levels increased by approximately 10% in both groups. There was no difference between the groups in bleeding patterns, with amenorrhea being the most frequent pattern in both groups (41.2% each) 12 months after LNG-IUS insertion. OAT did not increase the frequency of prolonged and/or frequent bleeding. Conclusion: OAT users and non-users had similar bleeding patterns after insertion of the LNG-IUS. Hemoglobin and hematocrit levels increased in both groups

Fundamento: A ativação da cascata inflamatória está presente na insuficiência cardíaca(IC). Existe relação entre esta ativação e estado protrombótico nesta síndrome. Dentre as etiologias de IC, a cardiomiopatia chagásica (CMC) parece ter maior ativação inflamatória e prognóstico mais reservado, possivelmente por especial risco para fenômenos tromboembólicos. A relação entre atividade inflamatória e protrombótica na cardiomiopatia chagásica e em outras etiologias é obscura. Objetivo: Estudar o perfil de marcadores protrombóticos e proinflamatórios em pacientes com insuficiência cardíaca chagásica comparando-os com os de etiologia não-chagásica. Métodos: Corte transversal. Critérios de inclusão: fração de ejeção do VE (FEVE) < 45% e tempo de início de sintomas > 1 mês. Os pacientes foram divididos em dois grupos: grupo 1(G1) sorologias positivas para Chagas e grupo 2(G2) sorologia negativa para Chagas. Dosou-se como fatores proinflamatórios: fator de necrose tumoral-alfa (TNF-), interleucina-6 (IL-6) e proteína C reativa (PCR) ultrassensível; fatores protrombóticos: dímero D, P-selectina solúvel, antígeno do fator de von Willebrand, fibrinogênio, complexo trombina-anti-trombina(TAT), fator tecidual(FT) e tromboelastograma(TEG). A amostra foi calculada para poder de 90%, assumindo-se diferença de 1/3 de desvio-padrão entre os grupos; p significativo se < 0,05. Análise estatística: teste exato de Fischer para comparação de proporções; teste t de student não-pareado para variáveis contínuas de distribuição normal e teste de Mann-Whitney para variáveis contínuas de distribuição assimétrica. Realizada análise de co-variância para ajuste de potenciais influências de co-variáveis. Resultados: Entre 16 de janeiro de 2008 e 08 de abril de 2009, 287 pacientes com IC crônica foram consecutivamente selecionados em nível ambulatorial ou de internação, sendo 138 no G1 e 149 no G2. O G1 apresentava maior porcentual de pacientes internados, de CF III/IV, PA sistólica mais baixa, maior freqüência de RHJ, ascite, menor fração de ejeção e níveis mais altos de BNP. Por outro lado, a prevalência de HAS, DM e DLP foi superior no G2. Dos marcadores proinflamatórios, o TNF- foi maior no G1, independentemente de outros fatores de gravidade(p<0,0001). A IL-6, apesar de maior no G1, sofreu maior influência de outras variáveis de gravidade do que da etiologia chagásica. Os níveis de PCR ultrassensível estavam elevados em ambos os grupos embora sem diferença entre eles. Dentre os fatores protrombóticos o dímero-D(p<0,0001), o fator de von Willebrand(p<0,0001) e a P-selectina(p=0,0262) foram mais altos no G1 que no G2. Os níveis de FT e TAT foram semelhantes. O fibrinogênio foi mais alto no G2 que no G1(p=0,0424), assim como os parâmetros do TEG - MA(p=0,0044), G(p=0,0022) e TG(p=0,001), embora todos estivessem dentro dos limites de referência na maioria dos pacientes em ambos os grupos. Na análise de co-variância apenas o dímero-D e a P-selectina mantiveram-se diferentes entre os grupos, sendo que os níveis de P-selectina estavam normais na maioria dos pacientes de ambos os grupos. Conclusões: A atividade proinflamatória esteve aumentada nos pacientes com IC chagásica e não-chagásica. A inflamação medida pelo TNF- foi independentemente maior entre chagásicos.Observou-se maior estado protrombótico entre chagásicos medido pelo dímero-D, independentemente de outros fatores de gravidade. / Background: Inflammatory cascade activation is present in heart failure (HF). This activation is closely related to a prothrombotic state in this syndrome. Among HF etiologies, Chagas cardiomyopathy (CCM) seems to have greater inflammatory activation and worse prognosis, possibly because of special risk for thromboembolic phenomena. The relation of inflammatory and prothrombotic activity between CCM and other HF etiologies remains unclear. Objective: To assess the profile of prothrombotic and proinflammatory markers in patients with chagasic in comparison to non-chagasic systolic heart failure. Methods: Cross sectional study. Inclusion criteria: LV ejection fraction (LVEF) < 45% and time of symptoms onset > 1 month. Patients were divided into two groups: group 1 (G1) positive Chagas serology and group 2 (G2) negative serology. Proinflammatory factors determined: tumor necrosis factor (TNF-), interleukin-6(IL-6) and ultrasensitive C-reactive protein (CRP); prothrombotic factors: D-dimer, soluble P-selectin, von Willebrand factor(vWF), fibrinogen, thrombin-anti-thrombin complex(TAT), tissue factor(TF) and thromboelastography(TEG). Sample was calculated for an 90% power, assuming a difference of 1 / 3 of the standard deviation; p significant if < 0.05. Statistical analysis: Fischer exact test for proportions, non-paired Students t test for parametric continuous variables and Mann-Whitney test for non-parametric continuous variables. Covariance analysis was performed to adjust for possible covariables influence on results. Results: From january 16th to april 8th 2009, 287 chronic HF patients were consecutively included, 138 in G1 and 149 in G2. G1 showed larger proportion of inpatients, higher III/IV functional class, lower systolic blood pressure, higher frequency of hepatojugular reflux, ascites, lower left ventricle ejection fraction and higher levels of B-type natriuretic peptide (BNP). On the other hand, G2 had higher proportion of hypertension, diabetes and hypercholesterolemia. Among proinflammatory markers, TNF- levels were higher in G1, independently of other prognosis variables (p<0,0001). Although IL-6 levels were higher in G1, there was greater influence of other prognosis variables than chagasic etiology itself. CRP levels were above reference values but there was no difference between G1 and G2. Among prothrombotic markers, D-dimer(p<0,0001), vWF(p<0,0001) and soluble P-selectin(p=0,0262) levels were higher in G1 than in G2. TF and TAT levels were similar in both groups. Fibrinogen levels were higher in G2 than in G1 (p = 0.0424), as well as TEG parameters MA(p=0,0044), G(p=0,0022) and TG(p=0,001), even though all of them were in normal reference range in most patients in both groups. D-dimer and soluble P-selectin kept different among groups in covariance analysis. However, soluble P-selectin levels were in normal reference range in both groups. Conclusions: Proinflammatory activity was increased in both groups. Inflammation measured by TNF- was independently greater among CCM patients. Greater prothrombotic state measured by D-dimer was observed in CCM patients independently of other prognosis variables.

Embora pacientes em terapia dupla antiplaquetária, composta de aspirina e um antagonista do receptor purinérgico P2Y12, tenham melhor prognóstico, eles ainda sofrem eventos trombóticos. Frequentemente, relaciona-se o nível de inibição plaquetária destes pacientes à ineficácia do tratamento. A reatividade plaquetária destes pacientes é uma função do nível do bloqueio do receptor P2Y12 e dos níveis de óxido nítrico e prostaciclina, importantes mediadores endoteliais, os quais agem em sinergia tripla aumentando o nível dos nucleotídeos cíclicos intraplaquetários mantendo a plaqueta em estado quiescente. Pacientes com disfunção endotelial têm menor disponibilidade destes mediadores e, consequentemente, menor inibição plaquetária in vivo. Propõe-se uma nova estratégia terapêutica composta de um inibidor do receptor purinérgico P2Y12, um ativador da enzima guanilato ciclase e um inibidor da fosfodiesterase, visando aumento dos nucleotídeos cíclicos, resultando em um forte efeito antiplaquetário localizado e tendo aplicação direta à pacientes com disfunção endotelial. / Disorders on the cardiovascular (CV) system are responsible for 31% of deaths worldwide. When inappropriately activated, platelets can cause myocardial infarction and stroke. Dual anti-platelet therapy (DAPT), composed of acetylsalicylic acid and a P2Y12 receptor blocker (clopidogrel, prasugrel, etc) is recommended for the prevention of recurrent CV events. Whilst DAPT is associated with an improvement in patient outcomes, thrombotic events do still occur. Platelet reactivity of patients under DAPT is a function of the level of P2Y12 receptor blockade and the levels of nitric oxide (NO) and prostacyclin (PGI2), two important endothelium-derived autacoids. Therefore, a new antithrombotic therapy is proposed using soluble guanylate cyclase (sGC) allosteric modulators independent of NO and phosphodiesterase (PDE) inhibitors. Based on the results observed, the combination of a sGC activator and a PDE inhibitor given at low doses, in the presence of P2Y12 receptor blocker, could produce enhanced and localized platelet inhibition.

La glycoprotéine VI (GP VI) des plaquettes sanguines humaines est le récepteur principal du collagène, composé le plus thrombogénique d'une paroi vasculaire lésée. Ainsi, GPVI est souvent considérée comme une cible de premier plan pour développer des tests diagnostiques ou des stratégies thérapeutiques innovantes, efficaces et sûres afin d'améliorer encore la prise en charge des accidents ischémiques. Les anticorps monoclonaux et leurs fragments actifs produits par ingénierie moléculaire constituent aujourd'hui une nouvelle classe de biomolécules en plein essor avec des propriétés bien adaptées à des applications thérapeutiques et diagnostiques. Notre groupe a produit plusieurs anticorps monoclonaux anti-GPVI par immunisation génique de souris. Ces anticorps ont une affinité élevé pour leur cible. Ils de distinguent les uns des autres par leur spécificité épitopique ainsi que par les effets engendrés par leur liaison à GPVI. Parmi ces anticorps, l'un présente un fort potentiel diagnostique parce qu'il reconnait les formes mono- et dimériques de GPVI, mais sa liaison aux plaquettes peut induire une activation ou la perte de GPVI. Un autre anticorps présente un fort potentiel thérapeutique parce que ses fragments actifs monovalents obtenus par papaïnolyse neutralisent l'interaction entre les plaquettes et le collagène, sans activer les plaquette. Cependant, l'origine xénogénique de cet anticorps est responsable d'une forte immunogénicité qui en interdit des applications en médecine humaine. Dans cette étude, nous avons conçus un fragments variable d'anticorps simple chaine (scFv) utile pour quantifier l'expression de la GPVI à la surface des plaquettes sanguines. Ce scFv a été reformaté de façon à lui insérer un motif de reconnaissance de la Protéine L (PpL) qui facilite sa détection et sa purification sans avoir recours à un peptide "drapeau". Nous avons également humanisé et créé plusieurs fragments d'anticorps recombinants monovalents inhibiteurs de l'interaction GPVI / collagène. Ces fragments d'anticorps présentent un potentiel thérapeutique élevé. / Human platelets glycoprotein VI (GPVI) is evidenced to be a platelet receptor of major importance in the occurrence of arterial thrombosis. Thus, it can be considered to be of great interest in diagnosis and therapeutic of atheriosclerotic diseases. Antibodies are powerful molecules which can be used in both diagnostic as well as for therapeutic purposes due to their unique characteristics. Monoclonal and recombinant antibodies have antigen restricted specificity, high affinity and can be used in various assays. Moreover, the good knowledge of their structure and molecular engineering facilities now allows the antibody modulation according to desired properties.Our group has already produced several monoclonal antibodies to human GPVI by gene gun immunization against the immunoadhesin hGPVI-Fc, which differ in fine epitopespecificity, affinity and other functional properties (Lecut et al. 2003). One, 3J24, with diagnostic potential while the other, 9O12, has a therapeutic potential because it blocks the binding of GPVI to collagen. Its Fab fragment has been extensively characterized in vitro,ex vivo and in vivo for its antithrombotic properties.Here, we designed and reshaped a single-chain antibody fragment (scFv) based on 3J24variable domains for the quantification of GPVI with diagnostic potential. We were also involved in the design, production and functional evaluation of humanized anti-GPVI recombinant antibody fragments (scFvs and Fabs) with therapeutic properties.

Thesis. 1977. Ph.D.--Massachusetts Institute of Technology. Dept. of Mechanical Engineering. / MICROFICHE COPY AVAILABLE IN ARCHIVES AND ENGINEERING. / Vita. / Bibliography: p. 200-205. / by Roger D. Kamm. / Ph.D.

This thesis combines five published research papers investigating methods of improving the safety and control of oral anticoagulation, with the use of computer assisted dosage and the standardisation of the International Normalised Ratio (INR). The INR is a conventional measurement derived from the time it takes blood of a patient to form a clot and is used to monitor the effects of widely used oral anticoagulants such as warfarin for the prevention of stroke and other related disorders. The first paper investigates whether the use of computer-assisted programs was as safe and effective as medical staff manual dosage in the prevention of bleeding or thrombotic complications during oral anticoagulant treatment. This was an international multi-centre randomised study conducted by the European Action on Anticoagulation (EAA) investigating the clinical benefit of two computer programs, PARMA 5 (Italy) and DAWN AC (UK). Composite clinical events were reduced by 7.6% using computer programs, though not achieving statistical significance (p=0.1), showing computer programs to be not dissimilar to medical staff dosage. The second paper recommends guidelines for screening safety and effectiveness of other marketed computer programs based on the results of the EAA study. A process for a candidate computer program to achieve non-inferiority relative to the medical staff dosage arm from the EAA study is explained. The third paper introduces a modified approach to the 'Direct INR' method for the standardisation of INR termed the 'Prothrombin Time/INR Line' (PT/INR). This was directly compared to the local International Sensitivity Index (ISI) calibration procedure originally approved by the World Health Organisation and later by the United States Food and Drug Administration (FDA). Using manually certified lyophilised plasmas tested by specialist centres, the PT/INR Line using a set of 5 calibrant plasmas to establish a fitted line to estimate local INR was shown to be as effective as the FDA procedure. The fourth paper investigates the PT/INR Line further by using simulated sets of calibrant plasmas across the therapeutic range of 2.0-4.5 INR and determining the PT/INR Line. Local INR of five validation plasmas, certified by 3 centres using the manual PT technique, was determined using the estimated PT/INR Lines and compared with local ISI calibration. Using 4 or 5 calibrant plasmas to determine the PT/INR Line was shown to be as accurate as local ISI calibrations for reliable local INR.The fifth and final paper assessed INR variability and control in oral anticoagulant therapy using a method termed the Variance Growth Rate (VGR), and compared its predictive ability of adverse events with the Time in Target INR range (TIR), the conventional method used in evaluating the quality of oral anticoagulant therapy. The VGR method was shown to be a better predictor of adverse bleeding or thrombotic episodes in the short term period prior to an event (3 and 6 months) compared with TIR.

Submitted by Suzana Dias (suzana.dias@famerp.br) on 2018-10-22T17:35:56Z No. of bitstreams: 1 LucianaVenturaCardoso_tese.pdf: 1870589 bytes, checksum: 7ea7e78e76f10c8a1f0c87d8a6c0a5a4 (MD5) / Made available in DSpace on 2018-10-22T17:35:56Z (GMT). No. of bitstreams: 1 LucianaVenturaCardoso_tese.pdf: 1870589 bytes, checksum: 7ea7e78e76f10c8a1f0c87d8a6c0a5a4 (MD5) Previous issue date: 2018-04-13 / Venous insufficiency is linked to longevity; an aggravating factor worldwidely, its main complication is venous ulcer. The boot of Unna is one of the forms of compression therapy. The bioimpedanciometry investigates accurately the patient's water balance. Objective: To evaluate the lower limb edema during the day comparing the use of the Unna’s boot and the conventional dressing through bioimpedanciometry. Material and Method: Fifteen legs with active wounds were evaluated, from September 2014 to December 2016. Randomization was drawing and the statistical analysis was performed using the paired t test, considering an alpha error of 5%. Results: The female representation was 100% of the total participants, the age ranged from 50 to 76 years, a mean age was 63 years. A significant difference between volume variations between morning and afternoon was found among those who did not wear the Unna’s boot. When compared the difference of edema with and without the Unna’s boot, it was detected that without the Unna’s boot the edema volume was higher. Conclusion: Unna’s boot showed better results than conventional dressing in relation to the reduction of lower limb edema during the day in patients with chronic venous ulcer. / A insuficiência venosa está ligada à longevidade; um agravo mundial, sua principal complicação é a úlcera venosa. A bota de Unna é uma das formas de terapia de compressão. A bioimpedânciometria investiga com acurácia o balanço hídrico do paciente. Objetivo: Avaliar o edema de membro inferior no transcorrer do dia comparando o uso da bota de Unna e o curativo convencional por meio da bioimpedânciometria. Material e Método: Foram avaliadas 15 pernas com feridas ativas, teve duração de setembro 2014 a dezembro de 2016. A randomização foi por sorteio e a análise estatística foi realizada utilizando-se o teste t pareado, considerando erro alfa de 5%. Resultados: A representatividade feminina foi de 100% do total de participantes, a idade variou de 50 a 76 anos, a média da idade foi 63 anos, com desvio padrão SD= 7,5. Detectou-se uma diferença significante entre as variações de volume entre o período da manhã e da tarde entre os que não usaram a bota de Unna. Quando comparada a diferença do edema com e sem bota de Unna, detectou-se que sem o uso da bota, o volume do edema foi maior. Conclusão: A bota de Unna mostrou-se superior ao curativo convencional na redução do edema de membro inferior no transcorrer do dia, dos pacientes com úlcera venosa crônica.

Over 26 million Americans suffer from pulmonary disease, resulting in more than 150,000 deaths annually. Lung transplantation remains the only definitive treatment for many patients, but has meager survival rates and only approximately 1,700 of the 2,200 patients added to the lung transplant wait list each year are transplanted. Extracorporeal gas exchangers have been used as an alternative to mechanical ventilation in acute respiratory failure and as a bridge to transplantation in chronic respiratory failure. Current gas exchangers are limited by their high resistance and low biocompatibility that lead to patient complications and device clot formation. Therefore, there exists a dire need for improved devices that can act as destination therapy. To accomplish the goal of destination therapy, this dissertation discusses three studies that were performed to pave the way. First, I examined clot formation and failure patterns of two common clinical devices (Maquet’s CardioHelp (CH) and Quadrox (Qx)) to further our understanding of their limitations with respect to long-term support. Overall, it was demonstrated that the Qx devices fail earlier and more frequently than CH devices and result in a significantly greater reduction in platelet count, and that a four-inlet approach is beneficial. Next, I determined the optimal sweep gas nitric oxide (NO) concentration that minimizes platelet binding and activation while ensuring that blood methemoglobin (metHb) concentrations increase less than 5%. Miniature artificial lungs were attached to rabbits in a pumped veno-venous configuration and run for 4 h with NO added to the sweep gases in concentrations of 0, 100, 250, and 500 ppm (n=8 ea.). 100 ppm significantly reduced the amount of platelet consumption (p < 0.05), reduced platelet activation as measured by soluble p-selectin (p < 0.05), and had negligible increases in metHb and will thus be used in future experiments. Last, I tested the Pulmonary Assist Device (PAD) which was designed for long term use as a bridge to transplantation and destination therapy. Benchtop experiments were performed that confirmed that it meets our design and performance goals. From here, we are equipped to commence with 30-day PAD testing in sheep.