INTRODUÇÃO: A circulação extracorpórea (CEC) é um fator etiológico importante para a lesão pulmonar, observada após cirurgia cardíaca. No entanto, o impacto da CEC na função mucociliar respiratória é desconhecido. O objetivo do estudo foi avaliar os efeitos imediatos da CEC sobre o sistema de transporte mucociliar. MÉTODOS: 22 porcos mestiços das raças Large White e Landrace com peso entre 33 a 47 kg alocados nos grupos controle (n = 10) e CEC (n = 12) completaram o estudo. As técnicas de anestesia e ventilação mecânica foram padronizadas. Após a indução da anestesia, foi realizada traqueostomia e uma amostra do tecido traqueal foi excisado (T0) em ambos os grupos. Todos os animais foram submetidos a toracotomia e CEC aorto-bicaval foi instalada no grupo CEC e mantida durante 90 minutos. Após o desmame da CEC (T90), uma segunda amostra do tecido traqueal foi obtida 180 minutos após a traqueostomia (T180). Amostras de muco foram coletadas na traquéia por meio de broncoscopia em T0, T90 e T180. Frequência de batimento ciliar (FBC) e transporte mucociliar in situ (TMC) foram estudados em epitélio traqueal ex vivo. As características do muco respiratório in vitro foram estudadas por transportabilidade ciliar no palato de rã (VTM), Transporte do muco respiratório in vitro pela tosse (TMT), Ângulo de contato (AC) e da viscosidade do muco por viscosímetro Cone-Plate (VM). RESULTADOS: A FBC diminuiu no grupo CEC (13,09 ± 1,91 Hz vs 11,06 ± 2,1 Hz, p <0,05), mas não no grupo controle (13,42 ± 0,96 Hz vs 12, 98 ± 2,84 Hz). No momento T90, a viscosidade aparente avaliado em 100 RPM estava aumentada no grupo CEC em relação ao controle. Não foram observadas diferenças significativas no TMC, VTM, TMT e AC. No grupo de CEC, foi percebida a perda do epitélio ciliado, edema submucoso e infiltração de células inflamatórias na avaliação histológica da traqueia. CONCLUSÃO: A CEC compromete agudamente o sistema de transporte mucociliar traqueal. Novos estudos são necessários para avaliar se esse comportamento tem implicações clínicas / BACKGROUND: Cardiopulmonary bypass (CPB) is an important etiologic factor for lung injury observed after cardiac surgery. However, the impact of CPB on respiratory mucociliary function is unknown. The objective of this study was to assess the immediate effects of CPB on mucociliary transport system. METHODS: Twenty-two mixed breed of Large White and Landrace pigs with weight between 33 to 47kg assigned to control (n=10) and CPB groups (n=12) completed the study. The techniques of anesthesia and mechanical ventilation were standardized. After anesthesia induction, tracheotomy was performed and a tracheal tissue sample was excised (T0) in both groups. All animals underwent thoracotomy and aorto-bicaval CPB was installed in CPB group and maintained during 90 minutes. After weaning from CPB (T90), a second tracheal tissue sample was obtained 180 minutes after tracheotomy (T180). Mucus samples were collected from the trachea using a bronchoscope at T0, T90 and T180. Ciliary beat frequency (CBF) and in situ mucociliary transport (MCT) were studied in ex vivo tracheal epithelium. In vitro respiratory mucus characteristics were studied by mucociliary transportability in frog palate (MT), Cough clearance (CC), Contact angle (CA) and the mucus viscosity by Cone-Plate viscometer (MV). RESULTS: CBF decreased in CPB group (13.09 ± 1.91 Hz vs. 11.06 ± 2.1 Hz, p < 0.05) but not in control group (13.42 ± 0.96 Hz vs. 12.98 ± 2.84 Hz). At T90 Apparent viscosity evaluated at 100 RPM increased in CPB group compared to control. No significant differences were observed in MCT, MT, CA and CC. In CPB group, it was observed loss of ciliated epithelia, submucosal edema and inflammatory cells infiltration in tracheal histology. CONCLUSION: CPB acutely compromise the tracheal mucociliary transport system. New studies are necessary to investigate if this behavior has any clinical implication

Orientadores: Cecília Amélia de Carvalho Zavaglia, Aron José Pazin de Andrade / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Mecânica / Made available in DSpace on 2018-08-16T12:14:14Z (GMT). No. of bitstreams: 1 Leme_Juliana_M.pdf: 6074962 bytes, checksum: bee519676aedd638a750cd66743a87f8 (MD5) Previous issue date: 2010 / Resumo: Um modelo de bomba de sangue para circulação extracorpórea (CEC) está sendo desenvolvida, esta denominada Bomba Espiral (BE). Essa bomba é composta por um fuso cônico com rosca de duas entradas, montado por dois rolamentos em um eixo de aço inoxidável, fixado a um cone externo. O movimento de rotação do fuso cônico é gerado por acoplamento magnético a um motor, através de dois imãs de seis polos. Esse trabalho tem como objetivo desenvolver protótipos com modificações estruturais, baseados em estudos prévios, realizados no Baylor College of Medicine em 1997, e realizar ensaios de desempenho hidrodinâmico, para determinar a melhor geometria, e ensaios para determinar o Índice Normalizado de Hemólise (INH). Foram realizados ensaios In Vitro, em um circuito de teste fechado e os dados de pressão, fluxo e rotação foram registrados. Com os dados obtidos, foram geradas curvas de desempenho hidrodinâmico (pressão x fluxo) em diferentes rotações, para cada protótipo os quais foram comparados entre si. O modelo da BE que apresentou o melhor desempenho possui características como imã maior, cone externo mais cônico e um rotor com uma profundidade de rosca maior, e este foi comparado com uma bomba utilizada como referência, Bio-Pump®. Foi verificado que a BE possui um melhor desempenho hidrodinâmico. Foi realizado o ensaio In Vitro para determinação do INH, calculado a partir da Hemoglobina Livre no Plasma, seguindo as diretrizes das normas ASTM F1841 e F1830. Os resultados obtidos de INH foram considerados excelentes para esta aplicação, INH = 0,0118927 ±0,0061 g/100L para a BE e INH = 0,007373± 0,004942341 g/100L para a Bio- Pump®. Estes resultados não apresentam diferenças estatisticamente significantes quando comparados. Portanto, o desenvolvimento da BE resultou em um modelo viável e competitivo, apresentando excelente desempenho hidrodinâmico e um ótimo INH. O próximo passo será a confecção dos moldes injeção em policarbonato, para a realização de novos ensaios de desempenho hidrodinâmico, determinação do INH, estudo de visualização de fluxo, ensaios In Vivo e, posteriormente, Avaliação Clínica / Abstract: A model of blood pump has been developed, the Spiral Pump. This pump has an internal rotor, a conically shaped fuse, containing threads with double entrance on its surface. This rotor is supported by two bearings assembled at a stainless steel shaft, fixed to the housing's base. The rotation of the conical fuse is generated by magnetic coupling to a motor, using two annular magnets with six poles. The main objective of this paper is to describe the development methodology of this new blood pump to be for Cardiopulmonary Bypass, during cardiac surgery. Based on previous studies, six different prototypes were developed and constructed. Hydrodynamic performance tests and hemolysis test were conducted. The hydrodynamic performance tests were made in vitro, using a closed loop circuit where total pressures head, flows and rotational speed were registered. Hydrodynamic performance curves were generated (pressure x flow) in different rotational speeds, for each prototypes. The prototype #6 showed best results and the preliminary hemolysis tests were conducted to determine the Normalized Index of Hemolysis (NIH), calculate through Plasma Free Hemoglobin (ASTM F1841 and F1830). NIH results were satisfactory for this application, NIH = 0,0118927 ±0,0061 g/100L, not showing significant different when compared to results from a reference pump (Bio-Pump®, Meditronic). The next step is to produce pumps made by polycarbonate injection to be able perform new hemolysis tests, in vivo tests and, subsequently, clinical evaluation / Mestrado / Materiais e Processos de Fabricação / Mestre em Engenharia Mecânica

A circulação extracorpórea (CEC) substitui as funções do coração e pulmões durante o tempo principal da cirurgia cardíaca, permitindo que o cirurgião acesse o interior do órgão por longo período de tempo e assegurando a oxigenação dos tecidos e eliminação dos seus produtos finais. Entretanto, tal técnica não é utilizada rotineiramente em medicina veterinária devido aos elevados custos e também em decorrência das alterações deletérias que promove no organismo dos animais. Visando uma melhor compreensão das complicações decorrentes da CEC, possibilitando amenizá-las, aventou-se o estudo da conexina 43 (Cx43), proteína presente no miocárdio e responsável pela formação das junções intercelulares do tipo gap. O presente estudo avaliou cinco cães indenes, SRD, não submetidos à CEC, procedendo-se a colheita de amostras correspondentes à região do septo interventricular, átrios direito e esquerdo, ventrículos direito e esquerdo e posterior análise da Cx43 através da imunofluorescência, western blot e RT-PCR. Avaliou também cinco cães indenes, SRD, submetidos à toracotomia intercostal direita convencional e posterior instituição da CEC. Amostras de miocárdio foram colhidas em três distintos momentos, quais sejam: T0 (antes da CEC), T1 (após duas horas de CEC) e T2 (após uma hora de desmame da CEC), para verificar as possíveis alterações na (Cx43) decorrentes da CEC, pela técnica de imunofluorescência. Ao final do procedimento experimental, os animais foram sacrificados. Os resultados mostraram que nos animais indenes não operados, a Cx43 apresenta localização em todas as regiões estudadas, não apresentando diferença significativa com relação à expressão nas diferentes regiões. Porém, nos animais submetidos à CEC, a referida conexina apresentou-se diminuída. Por fim, concluímos que em cães a CEC por duas horas com uma hora de circulação espontânea é técnica exeqüível, porém determina sérias alterações clínicas e induz a processos patológicos. / Cardiopulmonary bypass (CPB) replaces the heart and lung function during the main period of cardiac surgery, and it allows the access of the surgeon inside the organ for a long time, ensuring tissue oxygenation and the elimination of end products. However, this procedure isn´t used as a routine in veterinary medicine because of the high costs, and besides it causes damage in the animal organism. In order to promote a better comprehension of the complications caused by CPB, and perhaps to assuage them, this work was designed to study the connexin 43 (Cx43), a protein present in myocardium which forms intercellular communications like gap junctions. To do so five health mongrel dogs not submitted to CPB were evaluated and tissue samples were taken from specific locals as ventricular septal, right and left atrium, right and left ventricle, to perform Cx43 analyses by immunofluorescence, western blot and RT-PCR. Were also studied others five health mongrel dogs submitted to a conventional right intercostal thoracotomy for CPB procedure. Samples of myocardium were taken in three different moments: T0 (before CPB), T1 (after two hours of CPB) and T2 (after one hour the end of CPB) to check possible alterations in Cx43 caused by CPB using immunofluorescence technique. The euthanasia of the animals was performed at the end of the experimental period. The results shown that in health animals not operated, the Cx43 is located in every studied local and there is no significant difference in the protein expression areas. Nevertheless, the CPB technique reduced the connexin expressions in the operated animals. Finally, we may conclude that the cardiopulmonary bypass for two hours and a spontaneous perfusion for one hour is a feasible procedure in dogs, but it determines serious clinical alterations and may induce pathophisyological process.

As conexinas são proteínas essenciais e estão diretamente relacionadas à propagação do impulso elétrico no coração, à velocidade de condução bem como à gênese de muitas afecções cardíacas. Em face da circulação extracorpórea (CEC) ainda ser utilizada de forma inconsistente na medicina veterinária e devido aos poucos estudos observados na literatura sobre os efeitos provocados pelo emprego da CEC na expressão da conexina 43 (Cx43) no miocárdio, objetivou-se avaliá-la em 15 animais da espécie canina, distribuidos em três grupos (C, CEC-1 e CEC -2) sendo, respectivamente, antes de realizada a CEC, com 60 minutos após esta e 60 minutos de CEC seguida de 30 minutos de restauração da perfusão espontânea. Avaliou-se a Cx43 pelas técnicas de imunofluorescência, western blot e RT-PCR em tempo real no tecido muscular cardíaco de regiões correspondentes aos átrios direito (AD) e esquerdo (AE), ventrículos direito (VD) e esquerdo (VE) e septo transverso. Os resultados indicaram a presença da Cx43 em todas as regiões do miocárdio nos grupos C, CEC-1 e CEC-2. A expressão da Cx43 variou significativamente em CEC-2 no AE e VD em relação ao grupo C (p<0,05). A expressão gênica de Gja1 (gene da Cx43) não apresentou diferença significativa entre os grupos estudados. Em CEC-2 identificou-se a presença de vacuolização na túnica média de artérias de pequeno calibre do miocárdio. Concluiu-se também que a canulação da aorta bem como a instalação do circuito de CEC no modo aorto-bicaval constitui técnica exequível / Connexins are essential proteins that are directly associated with electrical impulse propagation and speed of propagation in the heart. They also play a major role in numerous heart conditions. The use of cardiopulmonary bypass (CPB) in veterinary medicine is inconsistent and few studies describe the effect of cardiopulmonary bypass on the expression of connexin 43 (Cx43) in the myocardium. The objective of this study was to evaluate myocardial expression of Cx43. Connexin 43 of 15 dogs was assessed at 3 moments: prior to CPB (Group C); 60 minutes after CPB (Group CPB1); and 60 minutes after CPB followed by 30 minutes of spontaneous perfusion (Group CPB2). Assessment of Cx43 included immunofluorescence, western blot and RT-PCR real time of heart tissue samples from the right atrium (RA), left atrium (LA), right ventricle (LV), right ventricle (RV) and transverse septum. Our results showed the presence of Cx43 in all 5 areas of the myocardium in groups C, CPB1 and CPB2. A significant variation on the expression of Cx43 was observed when CPB2 LA and CPB2 RV were compared to group C (p<0,05). Expression of Gja1 (gene for Cx43) did not vary significantly among the study groups. Group CBP2 presented vacuolation of the tunica media of small myocardial arteries. We conclude that cannulation of the aorta and aorto-bicaval setup of the CPB circuit is feasible technique

Introdução e Objetivos: A circulação extracorpórea (CEC) pode alterar a cinética de fármacos, inclusive dos antibióticos. O objetivo deste estudo foi avaliar influência da CEC sobre a farmacocinética da cefuroxima e verificar se o esquema posológico proposto: 1, 5g em bolus, seguido por três bolus de 750 mg 6/6 horas por 24 horas, mantém concentrações plasmáticas adequadas em pacientes submetidos à revascularização do miocárdio (RM). Método: Foi realizado estudo prospectivo observacional com grupo controle comparando 10 pacientes submetidos à RM com CEC e 9 pacientes submetidos à RM sem CEC (Registro Clincal trials: NCT0122882). Amostras sanguineas foram coletadas sequencialmente após cada dose de antibiótico e analisadas por meio do método de cromatografia líquida de alta pressão (HPLC). Análise de variância (ANOVA) foi utilizada para a comparação das concentrações plasmáticas e Log-rank para comparar as curvas que avaliaram o tempo, após a administração da cefuroxima, para que fossem atingidas concentrações abaixo de 16 g/mL (quatro vezes a MIC- mínima inhibitory concentration); considerando p< 0,05. Resultados: A CEC com tempo médio de 59,7 min 21,1 minutos não alterou a farmacocinética ou as concentrações plasmáticas da cefuroxima. O clearance médio dp (mL/ Kg/ min) e a mediana da concentração mínima (mg/ dL) do grupo RM com CEC versus RM sem CEC foram 1,7 0,7 versus 1,6 0,6 (p= 0,67) e 6,1 versus 5,7 (p= 0,77), respectivamente. Ambos os grupos apresentaram diminuição nas concentrações plasmáticas influenciadas somente pelo tempo, após cada bolus de cefuroxima (p< 0,001). Concentrações acima de quatro vezes a MIC foram mantidas por três horas, por todos os pacientes, porém, após seis horas do primeiro bolus a probabilidade de manutenção das mesmas concentrações foi de 0,2 para o grupo RM com CEC e de 0,44 para o grupo RM sem CEC, p= 0,867. Após os demais bolus concentrações abaixo de 16 g/mL foram atingidas antes de três horas. Conclusão: A CEC não influenciou as concentrações plasmáticas ou a farmacocinética da cefuroxima. Os resultados da farmacocinética devem ser considerados para a escolha de um melhor esquema posológico / Background and Objectives: Cardiopulmonary bypass (CPB) can alter the kinetic of drugs, including antibiotics. The aim of this study was evaluation of the CPB influence on the plasma concentrations and pharmacokinetics of cefuroxime and assess whether the dosing regimen 1.5 g dose, followed by 750 mg 6/6h for 24h is adequate for antibiotic prophylaxis. Methods: A prospective controlled observational study compared 10 patients undergoing surgery with CPB and 9 submitted to off-pump surgery, (Clinical trials identifier: NCT0122882). After each cefuroxime dose, blood samples were sequentially collected and analyzed using high-efficiency chromatography (HPLC). Plasma concentrations were compared using variance analysis and log-rank test was employed to evaluate the differences between curves that quantified the fraction of patients with a remaining plasma concentration above 16 mg/L within 6 h after each bolus; considering P < 0.05 significant. Results: After each cefuroxime bolus, both groups presented a significant decrease in plasma concentration over time (p< 0.001). Mean CPB time of 59.7 ± 21.1 min did not change cefuroxime pharmacokinetics or plasma concentrations. The mean clearance ± SD (mL/kg/min) and median of minimum concentration (mg/dL) of the CPB group versus the off-pump group were 1.7 ± 0.7 versus 1.6 ± 0.6 (p= 0.67) and 6.1 versus 5.7 (p= 0.77), respectively. Up to 3 h, but not after 6 h, following the first bolus, all patients had plasma concentrations above 16 mg/L (CPB group= 0.2 and off-pump group= 0.44, p=0.867). After another bolus, concentrations below 16 mg/dL were reached before 3 h. Conclusions: CPB does not influence cefuroxime plasma concentration, but pharmacokinetic data should be considered when choosing intervals between doses

Abstract Cardiac surgery with cardiopulmonary bypass induces the activation of systemic inflammatory response syndrome (SIRS) and results in at least some degree of global myocardial ischemia. Although these responses are usually short-lived, they may lead to serious complications and organ system failures. The present study evaluated the effects of high-dose glucose-insulin (1IU/kg/h) treatment (GIK) administered with the hyperinsulinemic normoglycemic clamp technique and a leukocyte-depleting filter on markers of systemic inflammatory response and myocardial ischemia-reperfusion injury in certain cardiac surgical risk groups. The study involved four prospective randomized controlled clinical trials and 119 patients. Cardioprotective effects were measured as myocardial enzyme release, recovery of contractile function and incidence of arrhythmias in all studies. The hemodynamic and metabolic effects of high-dose glucose-insulin treatment were evaluated in patients admitted for combined aortic valve (AS) and coronary surgery (40) and for urgent coronary surgery (39), and the latter study also involved proinflammatory cytokine and C-reactive protein analyses. The impacts of leukocyte filter on the expression of neutrophil adhesion molecules along with proinflammatory cytokines were evaluated in patients admitted for combined aortic valve (AS) and coronary surgery (20) and for solitary coronary surgery (20). The high-dose glucose-insulin treatment was associated with better preserved myocardial contractile function and less need for inotropic support after combined aortic valve and coronary surgery (I) and attenuation of postoperative CRP release after urgent coronary surgery (II). No effects on postoperative myocardial enzyme release (I, II) or on proinflammatory cytokine responses (II) were detected. The number of hypoglycemic events was low. The use of a leukocyte filter throughout the cardiopulmonary bypass period increased the neutrophil adhesion molecule CD11b expression in patients with both normal and prolonged CPB times and was associated with an enhanced proinflammatory cytokine response (III, IV). In conclusion, high-dose glucose-insulin treatment is safe, but requires strict control of blood glucose level. It reduces the need for inotropic support in patients with compromised cardiac status. The use of leukocyte filter leads to increased leukocyte activation and proinflammatory reaction.

Submitted in fulfillment of the requirements for the degree of Masters of Health Sciences in Clinical Technology, Durban University of Technology, Durban, South Africa, 2016. / Introduction: The immediate post-cardiopulmonary bypass (CPB) immune responses and organ injuries in immune- compromised patients remain poorly documented. We conducted a prospective clinical study to determine whether or not human immunodeficiency virus (HIV) seropositive patients generate higher acute systemic inflammatory response and suffer greater myocardial injury, compared to HIV seronegative patients. Methodology: Sixty-one consecutive patients i.e. Thirty HIV seropositive patients and Thirty-one seronegative, undergoing elective cardiac valve(s) replacement were enrolled, over a period of nine months from a single center hospital, after informed consent was acquired. The C-reactive protein (CRP) and Erythrocyte sedimentation rate (ESR) were used as biomarkers of acute inflammatory response, and cardiac troponin I (cTnI) as a biomarker for measuring postoperative myocardial injury. Single tests were measured preoperatively and postoperatively, in both groups, and these were compared and correlated to perioperative events and CPB parameters. Results: The mean age group was similar between the HIV seropositive and negative group (37.8 and 37.1 years, respectively). Preoperatively both groups had relatively equal CRP levels (p=0.388), ESR levels (p=0.817) and cTnI (p=0.489). The CPB events and durations were significantly different between the two groups, CPB duration (p=0.021). Other CPB events include, clamp aortic duration (p=0.026), CPB blood transfusion (p=0.013), CPB total urine output (p=0.035) and CPB peak lactate (p=0.040). Postoperatively we observed significant increased biomarkers level in both groups, with no significant difference between the groups: mean CRP (p=0.115), mean ESR (p=0.214) and cTnI (p=0.363). We observed a significant negative correlation between the mean change in CRP levels and mechanical ventilation (r=- 0.548, p=0.002) in the seropositive group, but not in the uninfected group (r=0.025, p=0.893). The correlation between the difference in CRP and ICU stay was not significant between in both group (r=-0.231, p=0.229 and r=0.25, p=0.975, respectively). A significant positive correlation between postoperative cTnI and the inotropic support duration (r=0.384, p=0.040) was seen in the seropositive groups, but not in the negative group (r=0.092, p=0.622). Furthermore we observed a significant drop in CD4 cells postoperatively (p=<0.001) in the HIV seropositive group. Antiretroviral treatment appeared to influence the degree of change in CD4 cells postoperatively. Conclusion: We conclude that HIV positive patients’ postoperative reactions to cardiac surgery supported by CPB are similar to those of HIV seronegetive patients. We further report non-paralleling correlations between the biomarkers and perioperative events; however these do not seem to affect the overall outcomes between the two groups. / M

Severe congenital heart disease (CHD) is diagnosed in the United States 147.4 times per 100,000 live births, excluding still births and abortions^1. With the advancement of diagnostic methods, prenatal care, and screening modalities, the total CHD birth prevalence has increased substantially^2. In turn, this increases the number of cardiac surgery cases. With the advancement of technology and cardiac surgery, smaller and younger patients are undergoing more complex cardiac procedures that involve cardiopulmonary bypass (CPB). Neonates and infants undergoing CPB are susceptible to adverse effects of CPB on the coagulation cascade due to their smaller weight and hematologic immaturity^3,4. In addition to these physiological issues in neonates and infants, CPB decreases circulating coagulation factors and anti- thrombin III levels to 50% and platelet counts to 70%^5, which can contribute to the post-operative bleeding.During CPB, neonates' and infants' coagulation factors become extremely diluted causing multiple coagulation defects^6. Optimizing the CPB circuit volume and the use of anti-fibrinolytic, packed red blood cells (pRBCs), platelets, cryoprecipitate, and ultrafiltration are among the most widely used methods in preserving and aiding coagulation factors^3,7-9. Another method of improving hemodilution-related coagulation dysfunction bleeding is by transfusing Fresh Frozen Plasma (FFP)^10. However, there are only a small number of articles focusing on the effect of FFP in post-operative bleeding in neonates and infants following complex cardiac surgery with CPB. I postulate that adding FFP during CPB will lower the possibility of patients to experience post-operational bleeding, thus, shortening their length of stay (LOS).

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20250号 / 医博第4209号 / 新制||医||1020(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 伊達 洋至, 教授 戸口田 淳也, 教授 開 祐司 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

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