A cirurgia para preservação do membro ou Limb-sparing é uma técnica reconstrutiva que envolve ressecção da neoplasia óssea em bloco, associada ou não à artrodese da articulação adjacente, com a fixação do enxerto ósseo com placa ou haste intramedular bloqueada. A utilização de autoenxerto desvitalizado tem muitas vantagens quando comparado aos aloimplantes, entre elas a diminuição da reação imunológica, ausência dos riscos de transmissão de doenças, adequada conformação anatômica e menor custo. Dentre os meios utilizados para desvitalização extracorpórea de autoenxerto cortical ósseo, destaca-se o nitrogênio líquido, por manter as propriedades biomecânicas do osso e promover a morte das células neoplásicas com apenas um ciclo de submersão. O objetivo do presente trabalho foi avaliar o nitrogênio líquido como método de desvitalização extracorpórea de autoenxerto ósseo cortical diafisário de tíbia em ovelhas. Foram utilizados 12 animais, divididos em dois grupos com seis cada, submetidos à osteotomia de um segmento ósseo de 7 cm, remoção do periósteo e medula óssea e fixação do enxerto com placa de compressão dinâmica estreita e 8 parafusos corticais de 4,5 mm. No grupo controle (GC), o enxerto foi reimplantado logo após sua remoção. No grupo nitrogênio líquido (GNL), o enxerto foi desvitalizado em nitrogênio líquido. Foram realizadas avaliações clínicas e radiográficas até o 180º dia de pós-operatório. A taxa global de incorporação das interfaces enxerto/osso receptor foi de 100% no GC, com um tempo médio de 75 dias, e de 91,67% no GNL, com tempo médio de 84,54 dias, não havendo diferença estatística entre os grupos. Com isso, é possível concluir que o nitrogênio líquido é um método adequado de desvitalização extracorpórea de autoenxertos ósseos corticais, já que não interfere na taxa e no tempo de incorporação das interfaces. / The Limb-sparing surgery is a reconstructive technique that involves the bone malignant tumor wide en bloc resection, associated or not with the adjacent articulation arthrodesis, having the bone graft fixation performed by a plate or interlocking nail. The use of devitalized autografts has many advantages when compared to allografts, as the decrease of the immunological reaction, the lack of spreading diseases risks, adequate anatomical conformation and lower costs. Within the utilized means to devitalize cortical bone autograft, the liquid nitrogen stands out for maintaining the biomechanical properties of the bone and to promote death of the tumor cells with just one submersion cycle. The purpose of this study was to evaluate the liquid nitrogen as an extracorporeal devitalization method of cortical diaphysis tibial sheep bone autograft. Twelve animals were used, divided into two groups of six each, submitted to osteotomy of a bone fragment of seven cm, removal of periosteum and bone marrow, with posterior autograft fixation using a narrow dynamic compression plate with eight cortical screws of 4,5 mm. On the control group (CG), the graft was reimplanted soon after removal. On the liquid nitrogen group (LNG), the graft was devitalized on liquid nitrogen. Clinical and radiographic evaluations were made until the 180th post-operative day. The global incorporation rate of the graft / host bone interfaces was a 100% at the CG, with an average time of 75 days, and 91.67% at the LNG with an average time of 84.54 days, having no statistical difference between the groups. It is possible to conclude that the liquid nitrogen is an adequate extracorporeal devitalization method of cortical bone autografts, since it has no interference on the rate and time of the incorporation surfaces.

Orientador: Desanka Dragosavac, Luis Alberto Magna / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-14T08:33:07Z (GMT). No. of bitstreams: 1 Antunes_Nilson.pdf: 1392711 bytes, checksum: 89e9cd241a2037104655c013c2724bfc (MD5) Previous issue date: 2009 / Resumo: Objetivo: Investigar a eficácia da ultrafiltração na remoção de mediadores inflamatórios liberados pela circulação extracorpórea e correlacionar ultrafiltração com alterações da função orgânica de acordo com o "Seqüential Organ Failure Assessment Score". Métodos: Quarenta pacientes foram incluídos e randomizados em dois grupos: "sem ultrafiltração" (n=20; Grupo I) e "ultrafiltração" (n=20; Grupo II). Complementos 3 e 4 ativados, interleucina 1beta, 6, 8 e fator de necrose tumoral alfa foram dosados antes da indução anestésica (T1), 5 minutos antes da circulação extracorpórea (T2), no líquido ultrafiltrado (T3), 30 minutos (T4), 6 (T5), 12 (T6), 24 (T7), 36 (T8) e 48 (T9) horas após término da circulação extracorpórea. "Sequential Organ Failure Assessment Score" foi avaliado nos tempos 1, 6 e 9. Significância estatística foi estabelecida com p = 0,05. Resultados: No líquido ultrafiltrado, apenas níveis de fator de necrose tumoral alfa foram detectados. Níveis de complemento 3 ativado, nos tempos 5 e 7, e complemento 4 ativado, nos tempos 5 e 6, foram significativamente elevados no grupo sem ultrafiltração, e níveis de interleucina 6 foram elevados no grupo ultrafiltrado, nos tempos 7 e 8. Interleucina 1beta, 8, fator de necrose tumoral alfa, e "Sequential Organ Failure Assessment Score" não tiveram diferenças significantes entre os grupos. Conclusões: Ultrafiltração retira significativamente fator de necrose tumoral alfa, mas isto não influencia nos níveis séricos desta citocina. Ultrafiltração com o tipo de filtro usado neste estudo não filtra outros mediadores inflamatórios estudados e não diminui a disfunção orgânica no pós-operatório. Deverá ser usada apenas para controle volêmico nos pacientes submetidos à circulação extracorpórea. / Abstract: Objective: To investigate the effectiveness of ultrafiltration in removing inflammatory mediators released by cardiopulmonary bypass and to correlate ultrafiltration with alterations in organic function according to the Sequential Organ Failure Assessment Score. Methods: Forty patients were included and randomized into two groups: "no ultrafiltration" (n=20; Group I) and "ultrafiltration" (n=20; Group II). Activated complement 3 and 4, interleukins 1beta, 6, 8 and tumor necrosis factor alfa were measured prior to anesthesia induction (Time 1), 5 minutes before cardiopulmonary bypass (Time 2), in the ultrafiltrated fluid (Time 3), 30 minutes (Time 4), and 6 (Time 5), 12 (Time 6), 24 (Time 7), 36 (Time 8) and 48 (Time 9) hours following cardiopulmonary bypass. Sequential Organ Failure Assessment Score was evaluated at Time 1, 6 and 9. Statistical significance was established at p = 0.05. Results: In the ultrafiltrated fluid, only tumor necrosis factor alfa levels were detected. Levels of activated complement 3 at Times 5 and 7 and activated complement 4 at Times 5 and 6 were significantly higher in the unfiltered Group, and levels of interleukin 6 were higher in the filtered Group at Times 7 and 8. Interleukins 1beta, 8, tumor necrosis factor alfa, and the Sequential Organ Failure Assessment score were not significantly different between the groups. Conclusions: Ultrafiltration significantly filtered tumor necrosis factor alfa but did not influences serum levels of this cytokine. Ultrafiltration with the type of filter used in this study had no effect in organic dysfunction and should be used only for volemic control in patients undergoing cardiopulmonary bypass. / Universidade Estadual de Campi / Pesquisa Experimental / Doutor em Cirurgia

Pneumonia is one of the main causes of death worldwide, specially of the elderly and the children under 5 years old. The traditional antibiotic-based therapy faces a crisis due to the increase in resistance and a lack of new molecules approved. Recently, our research group demonstrated the photodynamic inactivation of streptococcal pneumonia in vivo, a technique to which the development of resistance is described to be unlikely. This study proposed to investigate the applicability of nebulization as a delivery method for photosensitizers, in the hope to advance the research of the photodynamic inactivation of bacterial pneumonia. First, the critical attributes for nebulization (droplet size and delivery rate), the extent of nebulization, and the stability of three photosensitizers were stablished, and they were all found to be compatible with the technique. Then, the delivery was validated in an animal model using the most promising compound. It was possible to activate it using extracorporeal infrared light without causing acute lung or liver damage. In conclusion, nebulization presented itself as a promising tool for the delivery of photosensitizers to the respiratory tract. / A pneumonia é uma das principais causas de morte no mundo, sobretudo de idosos e crianças menores de cinco anos. A terapia tradicional, baseada em antibióticos, enfrenta uma crise diante do aumento da resistência e do número reduzido de novas moléculas que são aprovadas. Recentemente, este grupo de pesquisa demonstrou a inativação fotodinâmica da pneumonia pneumocócica in vivo, uma técnica para a qual o surgimento de resistência é descrito como pouco provável. Este estudo se propôs a investigar a aplicabilidade da nebulização como método de entrega de fotossensibilizadores, na esperança de avançar a pesquisa da inativação fotodinâmica da pneumonia bacteriana. Inicialmente, os atributos críticos da nebulização (tamanho de gotícula e taxa de entrega), a extensão da dose nebulizada, e a estabilidade de três fotossensibilizadores foram estabelecidas. Todos eles se mostraram compatíveis com a técnica. Então, a entrega foi validada em um modelo animal, utilizando o composto mais promissor. Foi possível ativá-lo usando luz infravermelha extracorpórea sem que houvesse dano agudo pulmonar ou hepático. Em conclusão, a nebulização se mostrou uma ferramenta promissora na entrega de fotossensibilizadores ao trato respiratório.

Orientadores: Eduardo Tavares Costa, Francisco Ubaldo Vieira Júnior / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de Computação / Made available in DSpace on 2018-08-19T00:09:22Z (GMT). No. of bitstreams: 1 MedeirosJunior_JohannesDantasde_M.pdf: 2215491 bytes, checksum: e35d132e2a61efa2d559a4dba08e356e (MD5) Previous issue date: 2011 / Resumo: Circulação extracorpórea é um processo no qual o sangue circula externamente ao corpo com o intuito de manter a oxigenação e o fluxo do sangue adequados durante a realização de determinados procedimentos cirúrgicos; para realizá-la é utilizado um conjunto de técnicas e equipamentos cuja função é substituir temporariamente o coração e os pulmões. Contudo, a circulação extracorpórea é percebida pelo organismo como um agente agressor e um dos principais parâmetros relacionados aos danos que ela provoca é a hemólise. O uso de bombas propulsoras é um dos causadores de hemólise em procedimentos envolvendo circulação extracorpórea e um dos principais tipos de bombas propulsoras é a bomba de roletes. A hemólise provocada pelo uso de bomba de roletes é devida principalmente ao grau de oclusão utilizado. Há dois métodos geralmente utilizados para calibração da bomba de roletes: o método de velocidade de queda e o método de calibração dinâmica. Nesse trabalho é proposto um dispositivo para auxiliar o perfusionista a ajustar bombas de roletes por um método menos oclusivo, a calibração dinâmica. O dispositivo é baseado em um microcontrolador PIC 18F4523 e é utilizado em conjunto com transdutores de pressão descartáveis normalmente utilizados em procedimentos cirúrgicos. Foram determinadas as curvas características de três transdutores de pressão na faixa de 0 a 1000 mmHg. Os resultados das calibrações realizadas com o protótipo desenvolvido foram comparados com os resultados obtidos com uma placa de aquisição de dados comercial. Dois, dos três transdutores, foram submetidos a testes de fadiga e um foi mantido como referência. Para validação do dispositivo, sendo utilizado para realizar calibração dinâmica, foram testados ajustes em 150, 250, 350, 450 e 500 mmHg. Em cada ajuste foram realizadas 8 medições simultâneas de pressão média de calibração dinâmica com o dispositivo e com uma placa de aquisição de dados comercial. Após a realização de todos os testes, os transdutores foram novamente caracterizados. As curvas características dos três transdutores obtidas na caracterização inicial mostraram igualdade estatística (p > 0,05). Após os testes de fadiga não foram observadas alterações nas respostas dos transdutores de pressão no início e após a realização dos testes, nos mesmos pontos de pressão (p > 0,05). As medidas de pressão de calibração dinâmica realizadas com o protótipo apresentaram igualdade estatística (p > 0,05) para toda a faixa de pressão testada, quando comparados com os respectivos resultados obtidos com a placa de aquisição de dados. Conclui-se que os transdutores utilizados atualmente em procedimentos cirúrgicos podem ser utilizados na calibração dinâmica sem perdas de características e que o dispositivo construído pode ser utilizado em ambiente operatório para ajustes de bombas de roletes pelo método de calibração dinâmica / Abstract: Cardiopulmonary bypass is a procedure in which blood circulates outside the body in order to maintain oxygenation and blood flow conditions during the performance of certain surgical procedures; for this procedure, it is used a set of equipments whose function is to temporarily replace the heart and lungs. However, the cardiopulmonary bypass is perceived by the body as an aggressor agent and one of the main parameters related to the damage it causes is hemolysis. The use of driving pumps is a cause of hemolysis in procedures involving cardiopulmonary bypass and one of the main types of driving pumps is the roller pump. Hemolysis caused by the use of roller pump is mainly due to the degree of roller occlusion used. There are two methods commonly used for adjusting the roller pump: the method of drop rate and the dynamic calibration method. In this work, we developed a device to assist the perfusionist adjust roller pumps by a less occlusive method, the dynamic calibration method. The device is based on a PIC 18F4523 microcontroller and is used in conjunction with disposable pressure transducers commonly used in surgical procedures. We have characterized three pressure transducers in the range of 0 to 1000 mmHg. Calibration results obtained with the prototype were compared with those obtained with a data acquisition board. Two of the three transducers were submitted to fatigue tests and one was kept as a reference. To validate the constructed device, being used to perform dynamic calibration, adjustments were at 150, 250, 350, 450 and 500 mmHg, in each set were performed eight simultaneous measurements of dynamic calibration mean pressure with the device and with a data acquisition board. After all tests were conducted, the transducers were re-characterized. The three characteristic curves obtained in the initial characterization showed statistically equivalence (p > 0.05) among themselves. After the fatigue tests no changes were observed in the responses of pressure transducers in the beginning and in the end of the tests, at the same pressure points (p > 0.05). Measurements of dynamic calibration pressure obtained with the prototype showed statistically equivalence (p > 0.05) throughout the tested pressure range, when compared with the results obtained with the data acquisition board. We conclude that the transducers currently used in surgical procedures may be used for dynamic calibration without losing their characteristics and that the constructed device can be used in surgery to adjust the roller pumps by the dynamic calibration method / Mestrado / Engenharia Biomedica / Mestre em Engenharia Elétrica

Background: Temporary mechanical circulatory support device without sternotomy has been highly advocated for severe cardiogenic shock patient but little is known when coupled with amniotic stem cell therapy. Case presentation: This case reports the first dual therapy of temporary left ventricular extracorporeal device CentriMag with distal banding technique and human amniotic stem cell injection for treating a severe refractory cardiogenic shock of an 68-year-old female patient. A minimally-invasive off-pump LVAD was established by draining from the left ventricle and returning to the right axillary artery with distal arterial banding to prevent right upper extremity hyperperfusion. Amniotic stem cells were injected intramyocardially at the left ventricular apex, lateral wall, inferior wall, and right subclavian vein. Conclusion: The concomitant use of the temporary minimally-invasive off-pump CentriMag placement and stem cell therapy not only provided an alternative to cardiopulmonary bypass and full-median sternotomy procedures but may have also synergistically enhanced myocardial reperfusion and regeneration.

Many complications associated with heart surgery are due to the negative effects of extracorporeal circulation (ECC). Some of these complications may be attributed to ECC-induced activation of inflammation and coagulation pathways. The inflammatory reaction may be caused by the interaction of blood components with air and the artificial surfaces of the ECC, from substances produced due to ischaemia-reperfusion injury of the heart and lungs, and from increased release of endotoxin from ischemic intestines. Staphylococcus aureus (S. aureus) infections are the leading cause of respiratory, skin and soft tissue, and bloodstream infections. Nitric oxide (NO) is a gaseous signaling molecule involved in many physiological and pathological processes. The role of NO in infection and inflammation is complex. NO may contribute to morbidity by acting as a vasodilator, myocardial depressant, and cytotoxic mediator. On the other hand, NO may have a salutary role through microvascular, cytoprotective, immunoregulatory, and antimicrobial properties. A simulated extracorporeal circulation (SECC) model is a closed circuit, including a roller pump, an oxygenator, a venous reservoir and polyvinyl chloride (PVC) tubing, where human blood is circulated. The SECC model allows studies of the blood and its components, without any influence from other organ systems. The aim of this work was to investigate NO effects during SECC and in S. aureus infection. Study I. Human blood was circulated through SECC during 3 hours, and leukocyte granule release was studied. Results indicated that NO addition during SECC is pro-inflammatory by stimulating leukocyte activation and granule release, and has no effect on oxygen free radical production and interleukin release. Study II. Investigating the effect of NO on S. aureus growth in whole blood during 180 min SECC, results showed a 6.2 fold growth in the presence of NO. Results indicated that by stimulating the expression of inducible lactate dehydrogenase, specific to S. aureus, NO may improve S. aureus resistance to oxidative stress, giving the pathogen a survival advantage. Study III. In an in vitro system of SECC, we measured glyceryl trinitrate (GTN) induced changes in leukocyte activation in whole blood caused by S. aureus infestation, as well as the effect of GTN on S. aureus growth. Results indicated that GTN does not affect S. aureus growth during SECC and has no effect on SECC-induced leukocyte activation. Study IV. Whole blood concentrations of selected leukocyte adhesion molecules, complement system components and myeloperoxidase  were measured in an in vitro system of SECC. Results indicated that SECC induces the increased expression of some leukocyte markers and that GTN addition significantly reduces the expression of some leukocyte activation markers.

A aprotinina, um inibidor de serinoproteinase ácido resistente de massa molar de 7 kDa, é utilizada como insumo ou medicamento. O objetivo principal deste trabalho foi purificar a aprotinina a partir de pulmão suíno. Três procedimentos foram utilizados. O primeiro procedimento utilizou a coluna de tripsina-agarose, o segundo procedimento utilizou a filtração tangencial e coluna de tripsina-Sepharose. O terceiro procedimento utilizou três cromatografias: filtração em gel, troca-iônica e afinidade (tripsina-agarose). A aprotinina suína foi purificada de pulmão utilizando o terceiro procedimento. A seqüência parcial do gene da aprotinina suína apresentou 74% de identidade com a seqüência do gene da aprotinina bovina. Outros dois inibidores de serinoproteinases ácido resistentes foram purificados, são eles: o fragmento ativo do segundo domínio do inibidor de leucoprotease secretada (SLPI), e um segundo inibidor de alta massa molecular, provavelmente bikunina. O protocolo de purificação utilizado neste trabalho recuperou 85mg de aprotinina suína por kg de pulmão. / Aprotinin, an acid stable serine proteinase inhibitor with a molecular mass of 7 kDa, is used as a reagent or drug. The purification of the aprotinin from porcine lungs was the main objective of this work. Three procedures were used. The first one utilized the trypsin-agarose column. The tangential ultra filtration and trypsin-Sepharose column were used in the second procedure. And finally, the gel filtration, ion-exchange and affinity chromatography were employed in the third procedure. The porcine lung aprotinin was purified using the third procedure. The partial sequence of the aprotinin gene was obtained and showed 74% of the identity with the aprotinin bovine gene sequence. Another two acid stable serine proteinase inhibitors were purified: the active fragment of the secretory leukoprotease inhibitor second domain, and one high molecular mass inhibitor, probably bikunina. The purification protocol used in this work recovered 85mg of the porcine aprotinin from kg of lung.

京都大学 / 0048 / 新制・課程博士 / 博士(社会健康医学) / 甲第22380号 / 社医博第102号 / 新制||社||医11(附属図書館) / 京都大学大学院医学研究科社会健康医学系専攻 / (主査)教授 中山 健夫, 教授 小池 薫, 教授 今中 雄一 / 学位規則第4条第1項該当 / Doctor of Public Health / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(社会健康医学) / 甲第22649号 / 社医博第109号 / 新制||社医||11(附属図書館) / 京都大学大学院医学研究科社会健康医学系専攻 / (主査)教授 中山 健夫, 教授 川上 浩司, 教授 伊達 洋至 / 学位規則第4条第1項該当 / Doctor of Public Health / Kyoto University / DFAM

Extracorporeal Membrane Oxygenation (ECMO) is a life-saving therapy usedfor support in critical heart and/or lung failure. Patient’s blood is pumped viaan artificial lung for oxygenation outside of the body. The circuit is composedof a blood pump, cannulae for drainage and reinfusion, a membrane lung,tubing and connectors. Its use is associated with thromboembolic complicationsand hemolytic damage. Detailed numerical studies of two blood pumps anda lighthouse tip drainage cannula were undertaken to characterize the flowstructures in different scenarios and their link to platelet activation. The pumpsimulations were modelled according to manufacturer’s proclaimed use but alsoin off-design conditions with flow rates used in adult and neonatal patients.Lagrangian Particle Tracking (LPT) was used to simulate the injection ofparticles similar in size to platelets to compute platelet activation state (PAS).The results indicated that low flow rates impacted PAS similarly to high flowrates due to increased residence time leading to prolonged exposure to shearstress despite the fact that shear per se was lower at low flow rate. Regardingthe cannula, the results showed that a flow pattern similar to a jet in crossflowdeveloped at the side holes. A parameter study was conducted to quantifydrainage characteristics in terms of flow rate distribution across the holes wheninput variables of flow rate, modelled fluid, and hematocrit were altered. Thefindings showed, across all the cases, that the most proximal hole row drainedthe largest fraction of fluid. The effects due to the non-Newtonian nature ofblood were confined to regions far from the cannula holes and the flow structuresshowed very limited dependence on the hematocrit. A scaling law was found tobridge the global drainage performance of fluid between water and blood. / <p>QC 210906</p>