published_or_final_version / abstract / Paediatrics and Adolescent Medicine / Master / Master of Philosophy

Adenocarcinoma pancreático é um tumor maligno com mau prognóstico, e por isso há necessidade de aperfeiçoamento ou criação de novas estratégias terapêuticas. A vacinação baseada em DCs é uma das abordagens mais promissoras, uma vez que as DCs são as células apresentadoras de antígenos (APCs) mais potentes e centrais para a indução e manutenção de uma resposta imune. Entretanto, em pacientes com câncer, a geração e a função de DCs podem ser deficientes, impondo um obstáculo para o sucesso de seu uso. Neste trabalho, nós descrevemos a geração in vitro de DCs a partir de células mononucleares do sangue periférico (PBMC) obtidas de pacientes ictéricos com câncer de pâncreas e, também, o efeito do plasma ictérico (PI) sobre as culturas de DCs de doadores saudáveis. PBMCs foram separadas do sangue obtido de 22 pacientes e 22 doadores saudáveis. Células aderentes foram cultivadas com GM-CSF e IL-4 (50ng/mL) por 7 dias. No 5o dia, TNF-<font face=\"symbol\">a (50ng/mL) foi adicionado para a maturação das DCs. Culturas foram realizadas em 10% PI ou plasma normal (PN). Células não aderentes foram coletadas no 7o dia, marcadas com anticorpos monoclonais anti-CD86, CD11c, CD14 e HLA-DR e analisados por citometria de fluxo. Células de pacientes, cultivadas em 10% de PI, quando comparadas com células de doadores saudáveis, cultivadas em 10% PN, apresentaram expressão reduzida (p<0,05) de CD86 e HLADR. É interessante observar que células geradas de PBMCs de pacientes não expressaram CD11c, diferente das células derivadas de doadores saudáveis. A presença de PI nas culturas dos doadores saudáveis causou uma significante diminuição na porcentagem de expressão de células HLA-DR+, CD11c+, CD86. Finalmente, quando PBMCs de pacientes foram cultivadas em PN, houve um aumento na expressão de HLA-DR e CD86 (p<0,05). Os ensaios de proliferação demonstraram também que as células de pacientes ictéricos tiveram capacidade aloestimuladora de linfócitos reduzida, quando comparada a de células de doadores saudáveis. Estes dados indicam uma alteração importante na capacidade das células de pacientes se diferenciarem em DCs in vitro, um fenômeno que parece depender tanto de fatores solúveis presentes no plasma e sobre as próprias células. / Pancreatic adenocarcinoma (PAdc) is an aggressive malignancy with poor prognosis, urging for improved or new therapeutic strategies. DC-based vaccination is one of such promising approaches. DC are the most potent antigen-presenting cells and central to the induction and maintenance of an immune response. However, in cancer patients DC generation and function may be deficient, imposing an obstacle to the success of their use. Here, we describe the in vitro generation of DC from peripheral blood mononuclear cells (PBMC) obtained from jaundiced patients with PAdc and, also, the effect of jaundiced plasma (JP) in the phenomenon. PBMC were separated from blood obtained from 10 patients and 10 healthy controls over a density gradient. Adherent cells were cultured with GM-CSF and IL-4 (50ng/mL) for 7 days. On the 5th day, TNF-<font face=\"symbol\">a (50ng/mL) was added for DC activation. Cultures were performed in 10% JP or normal plasma (NP). Non-adherent cells were harvested at day 7, labeled with FITC- or PE-conjugated monoclonal antibodies against CD86, CD11c, CD14, HLA-DR and analyzed by flow cytometry. Patients\' cells, cultured in 10% JP, compared to healthy donors\' cells, cultured in 10% NP, had a significantly (p<0.05) lower expression of CD86 and HLA-DR. It is noteworthy that cells generated from patients\' PBMC did not express CD11c, while from those derived from healthy donors\' cells did so. The presence of JP in healthy donors\' cells cultures caused a significant decrease in the percentage of HLA-DR+, CD11c+ and CD86+ cells. Finally, when patients\' PBMC were cultured in NP, a significant increase in HLA-DR and in CD86 expression occurred. MLR assays also demonstrated that cells from jaundice patients had decreased capacity to stimulate alloestimulation of lymphocytes when compared to healthy donors. These data indicate a significant alteration in the patients\' PBMC ability to differentiate into DC in vitro, a phenomenon that seems to depend both on soluble factors present in plasma and on the cells, themselves.

A coledocolitíase residual representa grande desafio na avaliação diagnóstica e proposta terapêutica. Neste contexto, realizamos estudo retrospectivo com o objetivo de avaliar critérios clínicos, laboratoriais e métodos de imagem para o seu diagnóstico; avaliar o resultado do tratamento através de procedimentos endoscópicos e cirúrgicos, bem como a ocorrência de complicações e sua repercussão no período de internação. Foram estudados 32 (trinta e dois) pacientes portadores de coledocolitíase residual internados na Clínica Cirúrgica do Hospital das Clínicas da Faculdade de Medicina da Universidade Federal de Goiás, no período de janeiro de 1995 a julho de 2001. Foram incluídos pacientes submetidos previamente a colecistectomia, nos quais o diagnóstico de coledocolitíase foi feito posteriormente (pela não realização de colangiografia trans-operatória) ou no próprio curso da colecistectomia, porém postergando-se o tratamento. Pudemos concluir: a maioria dos pacientes portadores de coledocolitíase residual tem como sintoma principal a icterícia; a ultra-sonografia não é um método diagnóstico eficaz, uma vez que demonstrou alteração de via biliar em cerca de 50% dos pacientes; tanto o procedimento endoscópico como o cirúrgico mostraram alto índice de clareamento da via biliar (acima de 90%), sendo a morbidade baixa e mortalidade nula em ambos os procedimentos; o período de internação foi menor quando o procedimento endoscópico foi realizado / Choledocholithiasis represents a great challenge in diagnostic evaluation and therapeutics. Because of it we\'ve proposed a retrospective study to analise the clinical and laboratorial criteria and image studies to the diagnosis of such condition. It was possible to evaluate the treatment (endoscopic or surgical) and complications with these information. Thus, with this aim, 32 (thirty-two) patients were evaluated. They were suffering from residual choledocholithiasis and were admitted on the Surgical Unit of the General Hospital of the Medical School of University of Goiás, from january 1995 to july 2001. It was included patients that were performed on a previous cholecistectomy. The diagnose of choledocholithiasis was get either during the surgery, although the definitive treatment had been postponed, or on the follow-up. We concluded that most part of the patients with residual choledocholithiasis were icteric and that the ultrasound study is not an effective method to detect residual choledocholithiasis (positive around 50%). Besides, both therapeutic procedures (endoscopic and surgical) were successful in the cleaning of biliary ducts (above 90%), had low morbidity and no mortality. The discharge of the patient was faster in case of endoscopic procedure

Introduction: Derangement of systemic and mucosal immunity, which are the integral components of the immune system, increases the risk of septic complications in patients postoperatively. The aims of this study were to investigate the integrity of systemic immunity as well as the mucosal immune system in patients with pancreatic cancer (PC), chronic pancreatitis (CP) and obstructive jaundice (OJ).Method: Healthy controls, as well as four groups of patients were studied. These included; jaundiced patients with PC, jaundiced patients secondary to benign disease (choledocholithiasis), non-jaundiced patients with PC and non-jaundiced patients with CP. The study evaluated the nutritional status including anthropometric measurements and the serum proteins: retinal binding protein (RBP), transferrin (TRF) and prealbumin (PALB). This study also evaluated systemic immunity in terms of total lymphocyte count, lymphocyte subsets (CD4+, CD8+, CD25+and CD56+), tumour necrosis factor alpha (TNF- alpha), interleukin-1alpha (IL-1 alpha) and complement components; and mucosal immunity in terms of CD3+, CD4+, CD8+, CD20+, CD57+, CD68+ and mast cells. Results: 78 patients were recruited (including 39 males) as follows: normal controls (n=17), benign OJ (n=9), patients with PC with jaundice (n=23), non-jaundiced patients with PC (n=20) and CP (n=9). Circulating CD25+ and CD4+ were significantly lower in the PC group whereas CD8+ showed increased levels in the same patients with a significant decrease in OJ patients when compared with controls. Circulating CD56+ showed no statistically significant difference between all four groups. In addition, IL-1 and TNF-alpha showed no statistically significant difference in all groups when compared with the control group. Also, C3 and CH50 showed significantly raised levels in PC with jaundice when compared with the control group. On the other hand mucosal lymphocyte subsets showed no statistically significant difference among all groups in comparison with the control group. As for prealbumin and transferrin, both showed significantly low levels in OJ, PC with jaundice and with PC when compared to healthy controls. Survival analysis for both PC groups was carried out and showed no difference in terms of age, however PC patients who survived over 13 months showed increased levels of prealbumin as well as low levels of CH50.Conclusion: Patients with PC both with and without jaundice showed some signs of altered and dysfunctional systemic immunity as well as a reduction in serum proteins. These findings may have implications on the disease progression and postoperative complications. This may warrant therapeutic interventions to restore nutrition and improve immunity before major surgical intervention is planned which could result in improving prognosis.

Thesis (M. Phil.)--University of Hong Kong, 2006. / Title proper from title frame. Also available in printed format.

No description available.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.

Adenocarcinoma pancreático é um tumor maligno com mau prognóstico, e por isso há necessidade de aperfeiçoamento ou criação de novas estratégias terapêuticas. A vacinação baseada em DCs é uma das abordagens mais promissoras, uma vez que as DCs são as células apresentadoras de antígenos (APCs) mais potentes e centrais para a indução e manutenção de uma resposta imune. Entretanto, em pacientes com câncer, a geração e a função de DCs podem ser deficientes, impondo um obstáculo para o sucesso de seu uso. Neste trabalho, nós descrevemos a geração in vitro de DCs a partir de células mononucleares do sangue periférico (PBMC) obtidas de pacientes ictéricos com câncer de pâncreas e, também, o efeito do plasma ictérico (PI) sobre as culturas de DCs de doadores saudáveis. PBMCs foram separadas do sangue obtido de 22 pacientes e 22 doadores saudáveis. Células aderentes foram cultivadas com GM-CSF e IL-4 (50ng/mL) por 7 dias. No 5o dia, TNF-<font face=\"symbol\">a (50ng/mL) foi adicionado para a maturação das DCs. Culturas foram realizadas em 10% PI ou plasma normal (PN). Células não aderentes foram coletadas no 7o dia, marcadas com anticorpos monoclonais anti-CD86, CD11c, CD14 e HLA-DR e analisados por citometria de fluxo. Células de pacientes, cultivadas em 10% de PI, quando comparadas com células de doadores saudáveis, cultivadas em 10% PN, apresentaram expressão reduzida (p<0,05) de CD86 e HLADR. É interessante observar que células geradas de PBMCs de pacientes não expressaram CD11c, diferente das células derivadas de doadores saudáveis. A presença de PI nas culturas dos doadores saudáveis causou uma significante diminuição na porcentagem de expressão de células HLA-DR+, CD11c+, CD86. Finalmente, quando PBMCs de pacientes foram cultivadas em PN, houve um aumento na expressão de HLA-DR e CD86 (p<0,05). Os ensaios de proliferação demonstraram também que as células de pacientes ictéricos tiveram capacidade aloestimuladora de linfócitos reduzida, quando comparada a de células de doadores saudáveis. Estes dados indicam uma alteração importante na capacidade das células de pacientes se diferenciarem em DCs in vitro, um fenômeno que parece depender tanto de fatores solúveis presentes no plasma e sobre as próprias células. / Pancreatic adenocarcinoma (PAdc) is an aggressive malignancy with poor prognosis, urging for improved or new therapeutic strategies. DC-based vaccination is one of such promising approaches. DC are the most potent antigen-presenting cells and central to the induction and maintenance of an immune response. However, in cancer patients DC generation and function may be deficient, imposing an obstacle to the success of their use. Here, we describe the in vitro generation of DC from peripheral blood mononuclear cells (PBMC) obtained from jaundiced patients with PAdc and, also, the effect of jaundiced plasma (JP) in the phenomenon. PBMC were separated from blood obtained from 10 patients and 10 healthy controls over a density gradient. Adherent cells were cultured with GM-CSF and IL-4 (50ng/mL) for 7 days. On the 5th day, TNF-<font face=\"symbol\">a (50ng/mL) was added for DC activation. Cultures were performed in 10% JP or normal plasma (NP). Non-adherent cells were harvested at day 7, labeled with FITC- or PE-conjugated monoclonal antibodies against CD86, CD11c, CD14, HLA-DR and analyzed by flow cytometry. Patients\' cells, cultured in 10% JP, compared to healthy donors\' cells, cultured in 10% NP, had a significantly (p<0.05) lower expression of CD86 and HLA-DR. It is noteworthy that cells generated from patients\' PBMC did not express CD11c, while from those derived from healthy donors\' cells did so. The presence of JP in healthy donors\' cells cultures caused a significant decrease in the percentage of HLA-DR+, CD11c+ and CD86+ cells. Finally, when patients\' PBMC were cultured in NP, a significant increase in HLA-DR and in CD86 expression occurred. MLR assays also demonstrated that cells from jaundice patients had decreased capacity to stimulate alloestimulation of lymphocytes when compared to healthy donors. These data indicate a significant alteration in the patients\' PBMC ability to differentiate into DC in vitro, a phenomenon that seems to depend both on soluble factors present in plasma and on the cells, themselves.

A coledocolitíase residual representa grande desafio na avaliação diagnóstica e proposta terapêutica. Neste contexto, realizamos estudo retrospectivo com o objetivo de avaliar critérios clínicos, laboratoriais e métodos de imagem para o seu diagnóstico; avaliar o resultado do tratamento através de procedimentos endoscópicos e cirúrgicos, bem como a ocorrência de complicações e sua repercussão no período de internação. Foram estudados 32 (trinta e dois) pacientes portadores de coledocolitíase residual internados na Clínica Cirúrgica do Hospital das Clínicas da Faculdade de Medicina da Universidade Federal de Goiás, no período de janeiro de 1995 a julho de 2001. Foram incluídos pacientes submetidos previamente a colecistectomia, nos quais o diagnóstico de coledocolitíase foi feito posteriormente (pela não realização de colangiografia trans-operatória) ou no próprio curso da colecistectomia, porém postergando-se o tratamento. Pudemos concluir: a maioria dos pacientes portadores de coledocolitíase residual tem como sintoma principal a icterícia; a ultra-sonografia não é um método diagnóstico eficaz, uma vez que demonstrou alteração de via biliar em cerca de 50% dos pacientes; tanto o procedimento endoscópico como o cirúrgico mostraram alto índice de clareamento da via biliar (acima de 90%), sendo a morbidade baixa e mortalidade nula em ambos os procedimentos; o período de internação foi menor quando o procedimento endoscópico foi realizado / Choledocholithiasis represents a great challenge in diagnostic evaluation and therapeutics. Because of it we\'ve proposed a retrospective study to analise the clinical and laboratorial criteria and image studies to the diagnosis of such condition. It was possible to evaluate the treatment (endoscopic or surgical) and complications with these information. Thus, with this aim, 32 (thirty-two) patients were evaluated. They were suffering from residual choledocholithiasis and were admitted on the Surgical Unit of the General Hospital of the Medical School of University of Goiás, from january 1995 to july 2001. It was included patients that were performed on a previous cholecistectomy. The diagnose of choledocholithiasis was get either during the surgery, although the definitive treatment had been postponed, or on the follow-up. We concluded that most part of the patients with residual choledocholithiasis were icteric and that the ultrasound study is not an effective method to detect residual choledocholithiasis (positive around 50%). Besides, both therapeutic procedures (endoscopic and surgical) were successful in the cleaning of biliary ducts (above 90%), had low morbidity and no mortality. The discharge of the patient was faster in case of endoscopic procedure

No description available.