Towards an advanced therapy medicinal product based on mesenchymal stromal cells isolated from the umbilical cord tissue: quality and safety data.

INTRODUCTION: Standardization of mesenchymal stromal cells (MSCs) manufacturing is urgently needed to enable translational activities and ultimately facilitate comparison of clinical trial results. In this work we describe the adaptation of a proprietary method for isolation of a specific umbilical cord tissue-derived population of MSCs, herein designated by its registered trademark as UCX®, towards the production of an advanced therapy medicinal product (ATMP). METHODS: The adaptation focused on different stages of production, from cell isolation steps to cell culturing and cryopreservation. The origin and quality of materials and reagents were considered and steps for avoiding microbiological and endotoxin contamination of the final cell product were implemented. Cell isolation efficiency, MSCs surface markers and genetic profiles, originating from the use of different medium supplements, were compared. The ATMP-compliant UCX® product was also cryopreserved avoiding the use of dimethyl sulfoxide, an added benefit for the use of these cells as an ATMP. Cells were analyzed for expansion capacity and longevity. The final cell product was further characterized by flow cytometry, differentiation potential, and tested for contaminants at various passages. Finally, genetic stability and immune properties were also analyzed. RESULTS: The isolation efficiency of UCX® was not affected by the introduction of clinical grade enzymes. Furthermore, isolation efficiencies and phenotype analyses revealed advantages in the use of human serum in cell culture as opposed to human platelet lysate. Initial decontamination of the tissue followed by the use of mycoplasma- and endotoxin-free materials and reagents in cell isolation and subsequent culture, enabled the removal of antibiotics during cell expansion. UCX®-ATMP maintained a significant expansion potential of 2.5 population doublings per week up to passage 15 (P15). They were also efficiently cryopreserved in a DMSO-free cryoprotectant medium with approximately 100% recovery and 98% viability post-thaw. Additionally, UCX®-ATMP were genetically stable upon expansion (up to P15) and maintained their immunomodulatory properties. CONCLUSIONS: We have successfully adapted a method to consistently isolate, expand and cryopreserve a well-characterized population of human umbilical cord tissue-derived MSCs (UCX®), in order to obtain a cell product that is compliant with cell therapy. Here, we present quality and safety data that support the use of the UCX® as an ATMP, according to existing international guidelines.

Use of intravascular ultrasonography for the characterization of coronary artery disease in patients with chronic kidney disease.

INTRODUCTION: Chronic kidney disease patients present a very high cardiovascular mortality. Nevertheless, a comparative description of lesion characteristics, using intravascular ultrasound in dialysis patients, has not yet been reported. The objective of the present study was to analyze the plaque morphology through intravascular ultrasound in comparison to their counterparts with normal renal function. METHODS: Patients were screened for coronary artery disease, and the coronary angiography was performed when indicated. Plaque morphology was evaluated by ultrasound, and findings were compared to a group of patients with coronary artery disease, who presented normal renal function, it carefully matched for all Framingham risk factors and lesion location at the coronary artery tree. RESULTS: One hundred and thirty-nine patients from a single center of hemodialysis were screened for the study. Patients with coronary lesions confirmed at the angiography presented lower hemoglobin (10.8 ± 1.5 versus 12.0 ± 19; p < 0.046) levels and higher levels of low-density lipoprotein (110.6 ± 25.8 versus 75.5 ± 43.1; p < 0.004), when compared to the ones without coronary artery disease. The ultrasound revealed greater proximal reference diameter (4.1 ± 0.6 versus 3.7 ± 0.5; p < 0.007), smaller crossed sectional area (4.2±1.6 versus 5.2 ± 1.8; p < 0.02), and the calcification was located in a deeper arterial layer (69 versus 9%; p < 0.004) in patients with chronic kidney disease when compared to the Control Group. CONCLUSION: Lesions of the patients with chronic kidney disease presented a larger proximal diameter and intense calcification in the deeper layer of the vessel, which suggest a greater positive remodeling effect in response to a more aggressive atherosclerotic process in the medial section of the artery.

Uso do ultrassom intracoronariano para a caracterização da doença arterial coronariana em pacientes com doença renal crônica / Use of intravascular ultrasonography for the characterization of coronary artery disease in patients with chronic kidney disease

INTRODUÇÃO: Neste estudo, objetivou-se identificar os pacientes em alto risco de desenvolvimento de DAC entre todos os indivíduos com DRC tratados em um grande centro universitário para estimar a prevalência de DAC e determinar a morfologia das placas ateroscleróticas através do IVUS em comparação com seus pares com função renal preservada. MÉTODOS: Pacientes foram investigados à procura de doença arterial coronariana, e a angiografia coronariana foi realizada quando indicada. Após avaliação sistemática para DAC os pacientes que apresentaram indicação foram encaminhados para cineangiocoronariografia e IVUS. As imagens de IVUS foram comparadas as de um grupo de pacientes com DAC, mas com função renal preservada, pareados cuidadosamente para todos os fatores analisados. RESULTADOS: Cento e trinta e nove pacientes de um centro de hemodiálise foram analisados para o estudo. Aqueles que tiveram a confirmação das lesões coronarianas na angiografia mostraram níveis séricos mais baixos de hemoglobina (10,8 ± 1,5 versus 12,0 ± 19; p < 0,046) e níveis elevados de LDL (110,6 ± 25,8 versus 75,5 ± 43,1; p < 0,004) quando comparados àqueles sem DAC. O IVUS revelou um maior diâmetro de referencia proximal e uma maior área de secção transversal nos pacientes com DRC em comparação ao grupo com função renal normal (4,1 ± 0,6 versus 3.7 ± 0,5; p < 0,007 e 4,2 ± 1,6 versus 5,2 ± 1,8; p < 0,02, respectivamente). Calcificação coronariana foi identificada pelo IVUS em 81 por cento das lesões no grupo com DRC, 31 por cento delas em mais de 180º da circunferência arterial. Os depósitos de cálcio estavam localizados em uma camada mais profunda da parede arterial nos pacientes com DRC (69 versus 30 por cento; p < 0,004). CONCLUSÕES: As lesões coronarianas apresentaram um maior diâmetro proximal e uma calcificação mais intensa em camada profunda da parede arterial sugerindo um efeito de remodelamento positivo exacerbado em resposta a um processo aterosclerótico mais agressivo na porção medial da parede arterial.

INTRODUCTION: Chronic kidney disease patients present a very high cardiovascular mortality. Nevertheless, a comparative description of lesion characteristics, using intravascular ultrasound in dialysis patients, has not yet been reported. The objective of the present study was to analyze the plaque morphology through intravascular ultrasound in comparison to their counterparts with normal renal function. METHODS: Patients were screened for coronary artery disease, and the coronary angiography was performed when indicated. Plaque morphology was evaluated by ultrasound, and findings were compared to a group of patients with coronary artery disease, who presented normal renal function, it carefully matched for all Framingham risk factors and lesion location at the coronary artery tree. RESULTS: One hundred and thirty-nine patients from a single center of hemodialysis were screened for the study. Patients with coronary lesions confirmed at the angiography presented lower hemoglobin (10.8 ± 1.5 versus 12.0 ± 19; p < 0.046) levels and higher levels of low-density lipoprotein (110.6 ± 25.8 versus 75.5 ± 43.1; p < 0.004), when compared to the ones without coronary artery disease. The ultrasound revealed greater proximal reference diameter (4.1 ± 0.6 versus 3.7 ± 0.5; p < 0.007), smaller crossed sectional area (4.2±1.6 versus 5.2 ± 1.8; p < 0.02), and the calcification was located in a deeper arterial layer (69 versus 9 percent; p < 0.004) in patients with chronic kidney disease when compared to the Control Group. CONCLUSION: Lesions of the patients with chronic kidney disease presented a larger proximal diameter and intense calcification in the deeper layer of the vessel, which suggest a greater positive remodeling effect in response to a more aggressive atherosclerotic process in the medial section of the artery.

Increased calcification and protein nitration in arteries of chronic kidney disease patients.

BACKGROUND: Cardiovascular disease in chronic kidney disease (CKD) has peculiar characteristics. The aim of this study was to analyze atherosclerosis, vascular calcification and nitration in arteries from CKD patients. METHODS: External iliac and renal artery segments from 27 stage 5 CKD patients and 25 donor controls, respectively, were collected during the transplantation procedure. RESULTS: CKD patients presented a significantly higher degree of lesion. In a large proportion (72%) of CKD patients, we observed vascular calcifications. Immunohistochemistry for nitrotyrosine revealed a significant increase in nitrotyrosine production in arteries from CKD patients compared with control donors. In addition, within CKD patients, nitrotyrosine staining was significantly stronger in arteries with media calcification when compared with arteries without media calcification. CONCLUSION: The arteriopathy in the CKD patients appears in an early age and seems to be distinct from the arteriopathy of the general population, especially due to intense calcification and vascular oxidative stress.

Avaliação e manejo da doença cardiovascular em pacientes com doença renal crônica / Assessment and management of cardiovascular disease in patients with chronic renal disease

Doenças cardiovasculares são as principais complicações fatais da doença renal crônica, tanto para pacientes em terapia renal substitutiva quanto para aqueles em tratamento conservador. A análise de seus fatores de risco, abordagem diagnóstica e adequado tratamento são fundamentais para a redução de mortalidade associada a essas complicações. Neste artigo de revisão são discutidos aspectos de fisiopatologia, métodos de investigação e abordagem terapêutica da doença cardiovascular na doença renal crônica.

Cardiovascular diseases are the major fatal complications of chronic kidney disease, both for patients in renal replacement therapy and for those on dialysis. The analysis of their risk factors, diagnosis and appropriate treatment are key to reducing mortality associated with these complications. In this review we discuss aspects of pathophysiology, research methods and therapeutic approach to cardiovascular disease in chronic kidney disease.

[Assessment and management of cardiovascular disease in patients with chronic kidney disease].

Cardiovascular disease is the leading cause of death in the set of chronic kidney disease (CKD) patients, whether on renal replacement therapy or conservative treatment. A better understanding of cardiovascular risk factors, diagnostic approach and management are central keys to develop strategies to reduce cardiovascular mortality among those patients. This review article discusses some aspects of pathophysiology, investigation methods and current treatment of cardiovascular disease in CKD patients.

Analysis of the immediate outcomes of a comparative randomized study between aorto-saphenous mechanical anastomosis versus conventional anastomosis.

OBJECTIVE: To evaluate the immediate results of mechanical aorto-saphenous anastomosis compared with conventional anastomosis. METHOD: We evaluated 12 patients. The mean age ranged from 62.33 +/- 7.30 years. Of 12 patients submitted to surgery without extracorporeal circulation, 10 (83.3%) patients were male. Thirty-three proximal anastomoses were evaluated, 21 of them being performed through the conventional manner and 12 with the St. Jude Symmetry aortic connector. The time spent on anastomosis, and free flow and patency on the 4th day postoperative were analysed. RESULTS: The mechanical anastomosis was successfully performed in all patients. Electrocardiographic alteration compatible with myocardial infarctation (MI) on the 2nd day postoperative was observed only in one patient. The patient was referred to angiographic restudy, becoming evident a conventional proximal anastomosis occlusion for the marginal branch. Three patients had atrial fibrillation. The average time spent to perform the mechanical anastomosis was 44.08 +/- 9.26 seconds against 3.86 +/- 0.61 minutes of the conventional anastomosis (p = 0.0022). The average blood free flow observed in the mechanical anastomosis was 302.75 +/- 82.76 mL/min versus 190.75 +/- 51.53 mL/min (p = 0.0022). In the angiographic restudy performed on the 4th postoperative day, it was detected the occlusin of three mechanical anastomosis. There was no new conventional anastomosis (p = 0.2500). CONCLUSION: The present study showed a statistically significant superiority for mechanical anastomosis of the saphenous vein with the aorta when evaluated the blood free flow and the time to perform the anastomosis. In relation to the artery condition in the postoperative angiography, one cannot say there was statistically significant difference between the procedures studied.

OPCAB in patients on hemodialysis.

OBJECTIVE: To analyze the hospital outcomes of patients, with chronic renal insufficiency in the hemodialysis, submitted to OPCAB. METHOD: Fifty-one patients with chronic renal insufficiency were submitted to OPCAB. Hemodialysis was performed on the day before and the day after the operation. Myocardial revascularization was performed using LIMA's suture and suction stabilization. RESULTS: Fifty-one patients, with an average of 61.28+/-11.09 years, were analyzed. Thirty patients (58.8%) were female. The predominant functional class was IV in 21 (41.1%) of the patients. The left ventricle ejection fraction was dire in 21 (41.1%) patients. The mean EUROSCORE of this series was 7.65+/-3.83 and the mean number of distal anastomosis was 3.1+/-0.78 per patient. The average time of mechanical ventilation was 3.78+/-4.35 hours and the mean ICU stay was 41.9+/-13.8 hours, while the average hospitalization was 6.5+/-1.31 days. In respect to complications, nine (17.6%) of the patients developed atrial fibrilation, and one (1.9%) patient presented with a case of ischemic stroke but had a good recovery during hospitalization. There were no deaths in this series. CONCLUSION: Chronic renal patients submitted to hemodialysis were always a high risk population for myocardial revascularization. In this series, the absence of extracorporeal circulation appeared to be safe and efficient in this special subgroup of patients. The operations were performed with low indices of complications, absence of deaths and relatively low stays in the ICU and in hospital.

Análise dos resultados imediatos de estudo comparativo entre anastomose mecânica aorto-safena versus convencional / Analysis of the immediate outcomes of a comparative randomized study between aorto-saphenous mechanical anastomosis versus conventional anastomosis

Objetivo: Avaliar os resultados imediatos das anastomoses mecânicas aorto-safena, comparando-as às anastomoses convencionais. Método: Foram analisados 12 pacientes, com idade média...

Revascularização do miocárdio sem circulação extracorpórea em pacientes submetidos à hemodiálise / OPCAB in patients on hemodialysis

OBJETIVO: Analisar evolução hospitalar dos pacientes portadores de insuficiência renal crônica (IRC) em hemodiálise, submetidos a operação sem circulação extracorpórea (CEC). MÉTODO: Cinqüenta e um pacientes portadores de IRC foram submetidos à operação sem CEC. A hemodiálise foi realizada no dia anterior à operação e no dia seguinte. A revascularização do miocárdio foi realizada com ponto de LIMA e estabilizador por sucção. RESULTADOS: A idade média foi de 61,28±11,09 anos e 31 (58,8 por cento) pacientes eram do sexo feminino. A classe funcional predominante foi a IV em 21 (41,1 por cento) dos pacientes. A fração de ejeção do ventrículo esquerdo era ruim em 21 (41,1 por cento) pacientes. O EUROSCORE médio desta série de pacientes foi de 7,65±3,83. O número médio de artérias coronárias revascularizadas foi de 3,1±0,78 por paciente. O tempo médio de ventilação mecânica foi de 3,78±4,35 horas. A permanência média na CTI foi de 41,9±13,8 horas, enquanto a média de permanência hospitalar foi de 6,5±1,31 dias. Quanto às complicações, nove (17,6 por cento) pacientes desenvolveram FA e um (1,9 por cento) apresentou quadro de AVC isquêmico, com boa recuperação durante a internação. Não houve óbito nesta série. CONCLUSÃO: Pacientes renais crônicos submetidos à hemodiálise sempre foram uma população de alto risco para revascularização do miocárdio. A ausência de CEC, aparentemente, cursa com baixos índices de morbi-mortalidade nesta população.

OBJECTIVE: To analyze the hospital outcomes of patients, with chronic renal insufficiency in the hemodialysis, submitted to OPCAB. METHOD: Fifty-one patients with chronic renal insufficiency were submitted to OPCAB. Hemodialysis was performed on the day before and the day after the operation. Myocardial revascularization was performed using LIMA's suture and suction stabilization. RESULTS: Fifty-one patients, with an averageof 61.28±11.09 years, were analyzed. Thirty patients (58.8 percent) were female. The predominant functional class was IV in 21 (41.1 percent) of the patients. The left ventricle ejection fraction was dire in 21 (41.1 percent) patients. The mean EUROSCORE of this series was 7.65±3.83 and the mean number of distal anastomosis was 3.1±0.78 per patient. The average time of mechanical ventilation was 3.78±4.35 hours and the mean ICU stay was 41.9±13.8 hours, while the average hospitalization was 6.5±1.31 days. In respect to complications, nine (17.6 percent) of the patients developed atrial fibrilation, and one (1.9 percent) patient presented with a case of ischemic stroke but had a good recovery during hospitalization. There were no deaths in this series. CONCLUSION: Chronic renal patients submitted to hemodialysis were always a high risk population for myocardial revascularization. In this series, the absence of extracorporeal circulation appeared to be safe and efficient in this special subgroup of patients. The operations were performed with low indices of complications, absence of deaths and relatively low stays in the ICU and in hospital.

[Surgical treatment without extracorporeal circulation of artery disease related to Takayasu's arteritis involving the aorta and great vessels]. / Tratamento cirúrgico sem circulação extracorpórea de arteriopatia relacionada com arterite de Takayasu envolvendo aorta e vasos da base.

We report the cases of 2 female patients with Takayasu's arteritis referred to our service with lesions affecting the descending thoracic aorta and great vessels. One of the patients had a critical obstructive lesion in the left coronary ostium. Both patients underwent surgery without extracorporeal circulation, with full heparinization and autotransfusion.

Tratamento cirúrgico sem circulacão extracorpórea de arteriopatia relacionada com arterite de Takayasu envolvendo aorta e vasos da base / Surgical treatment without extracorporeal circulation of artery disease related to Takayasu's arteritis involving the aorta and great vessels

Relatamos o caso de duas pacientes portadoras de arterite de Takayasu, encaminhadas ao nosso servico onde as lesões acometiam a aorta torácica descendente e os vasos da base e, em uma delas, lesão obstrutiva crítica do óstio da coronária esquerda. Ambas as pacientes foram operadas sem circulacão extracorpórea, com heparinizacão plena e auxílio de autotransfusão.

Revascularização total do miocárdio sem circulação extracorpórea: cinco anos de experiência / Total myocardium revascularization without extracorporeal circulation: five-year experience

Objetivo: Avaliar os resultados imediatos da operação para revascularização do miocárdio sem circulação extracorpórea, analisando-se complicações e mortalidade.Método: Foram submetidos à operação para revascularização do miocárdio sem circulação extracorpórea 1440 pacientes. A técnica operatória consistiu em oclusão proximal da artéria abordada, aplicação do ponto de LIMA na deflexão pericárdica posterior e estabilização da artéria alvo com estabilizador de sucção. As anastomoses distais foram feitas inicialmente.Resultados: Entre os pacientes avaliados, 924 eram ao sexo masculino, com idade média de 63,12 anos. A fração de ejeção era normal em 749 pacientes. Setecentos e quarenta(51,4 por cento) pacientes tinham antecedente de infarto do miocárdio. Seiscentos e oitenta e sete(47,6 por cento) dos pacientes encontravam-se em classe funcional III ou IV. O EuroSCORE médio foi de 4,93. A média de anastomoses distais foi de 3,12 por paciente. Mil cento e setenta e três(81,5 por cento) dos pacientes permaneceram menos de 12 horas em ventilação mecânica, sendo que destes, 888(61,7 por cento) permaneceram menos de 6 horas entubados. A permanência em UTI foi de uma noite em 330(22,8 por cento) dos pacientes e de duas noites em 930(64,6 por cento). Cento e oitenta e dois(12,4 por cento) pacientes permaneceram três ou mais noites na UTI. Quanto as complicações, três(0,2 por cento)pacientes apresentaram insuficiência renal, seis(0,4 por cento) tiveream acidente vascular cerebral, 19(1,3 por cento) foram reoperados por sangramento, 19(1,3 por cento) tiveram mediastinite, 18(1,25 por cento) infarto agudo do miocárdio e 212(14.7 por cento) apresentaram fibrilação atrial. Houve 50(3,5 por cento)óbitos, sendo 29(2,5 por cento)entre 1148 pacientes operados eletivamente, nove(4,7 por cento)entre 190 pacientes submetidos a reoperação coronariana e 12(11,7 por cento)entre 102 pacientes operados em caráter de emergência.Conclusão: Com a evolução da tecnologia biomédica, todos os vasos do coração passaram a ser abordados. Estes dados sugerem que a operação para a revascularização do miocárdio é segura e eficaz, podendo ser aplicada em todos os pacientes que necessitem de cirurgia coronariana com baixos índices de complicações e mortalidade

Revascularização do miocárdio sem circulação extracorpórea em pacientes acima de 75 anos: análise dos resultados imediatos / Myocardial revascularization without extracorporeal circulation in patients over 75 years of age: analysis of immediate results

OBJETIVO: Analisar os resultados imediatos da operação para revascularização do miocárdio sem circulação extracorpórea em pacientes acima de 75 anos. MÉTODOS: De janeiro 2001 a dezembro 2003, 193 pacientes com idade variando de 75 a 94 anos, correspondendo a 100 por cento das operações coronarianas realizadas em pacientes acima de 75 anos, sendo 121 homens e 72 mulheres, foram submetidos a operação para revascularização do miocárdio sem circulação extracorpórea e analisados, retrospectivamente, os dados de internação dos pacientes. RESULTADOS: A hipertensão arterial esteve presente em 174 (90 por cento), seguida de dislipidemia em 115 (59 por cento), tabagismo em 89 (46 por cento) e diabetes em 57 (29 por cento), e 96 (49,7 por cento) com antecedente de infarto do miocárdio. Destes, 53 (27,4 por cento) tinham apresentado evento isquêmico agudo em um período < 30 dias da data da operação. Tinham lesões obstrutivas críticas em três ou mais vasos coronarianos 156 (80,95 por cento) pacientes, enquanto 30 (15 por cento) apresentavam obstrução do tronco da coronária esquerda e 30 (15 por cento) apresentavam fração de ejeção < 30 por cento. A pontuação obtida através do EUROSCORE variou de 3 a 18, com uma mortalidade esperada para o grupo de 7,54±2,69 por cento. O número total de anastomoses distais foi de 639. Houve 7 (3,62 por cento) óbitos, três por broncopneumonia, um por mediastinite, um por causas metabólicas, uma morte súbita após refixação de deiscência de esterno e um choque cardiogênico. Reoperação por sangramento, reintubação orotraqueal e ventilação mecânica prolongada foram fatores associados com aumento da mortalidade. CONCLUSÃO: A operação adotada para revascularização do miocárdio sem circulação extracorpórea foi associada à baixa mortalidade, baixo índice de complicações e curto período de hospitalização.

The Wada test with propofol in a patient with epilepsy

The usual drug used in the Wada test is amobarbital, but it is not available in Brazil. Propofol was already used by Bazin et al. in 1998, and in their report the test resulted good in the absence of any adverse effect. We report the use of propofol as the anesthetic for the Wada test. The test was carried out in a 26 years old woman with temporal medial lobe epilepsy refractory to medical treatment. Language functions and memory were tested after injection in both hemispheres by three procedures (Seattle, Montreal and Interview procedures). Propofol showed to be good to carry on the Wada test.