ABSTRACT Purpose: Gene expressions of vascular Endothelial Growth Factor Alpha (VEGFa), Nuclear Factor Kappa-Light-Chain-Enhancer of Activated B cells (NFkB) and cytokines could be useful for identifying potential therapeutic targets to alleviate ischemia-reperfusion injury after liver transplantation. Cytokine gene expressions, VEGFa and NFkB were investigated in a preclinical swine model of liver transplantation. Methods: A total of 12 pigs were used as donors and recipients in liver transplantation without venovenous bypass or aortic clamping. NFkB, IL-6, IL-10, VEGFa and Notch1 gene expression were assessed. These samples were collected in two specific times: group 1 (n= 6) - control, samples were collected before recipient’s total hepatectomy and group 2 - liver transplantation group (n=6), where the samples were collected one hour after graft reperfusion. Results: Liver transplantation was successfully performed in all recipients. Liver enzymes were elevated in the transplantation group. NFkB gene expression was significantly decreased in the transplantation group in comparison with the control group (0.62±0.19 versus 0.39±0.08; p= 0.016). No difference was observed between groups Interleucine 6 (IL-6), interleucine 10 (IL-10), VEGFa and Notch homolog 1 (Notch1). Conclusions: In this survey a decreased NFkB gene expression in a porcine model of liver transplantation was observed. Purpose VEGFa, , (VEGFa) KappaLightChainEnhancer Kappa Light Chain Enhancer (NFkB ischemiareperfusion ischemia reperfusion Methods clamping IL6, IL6 IL 6, IL-6 IL10, IL10 10, IL-10 assessed times n= n (n recipient s n=6, n6 n=6 (n=6) Results 0.62±0.19 062019 0 62 19 (0.62±0.1 0.39±0.08 039008 39 08 p 0.016. 0016 0.016 . 016 0.016) (IL-6) (IL-10) Notch1. (Notch1) Conclusions (VEGFa IL- IL1 IL-1 (n=6 0.62±0.1 06201 (0.62±0. 0.39±0.0 03900 3 001 0.01 01 (IL-6 (IL-10 (Notch1 0.62±0. 0620 (0.62±0 0.39±0. 0390 00 0.0 (IL- (IL-1 (Notch 0.62±0 062 (0.62± 0.39±0 039 0. (IL 0.62± 06 (0.62 0.39± 03 0.62 (0.6 0.39 0.6 (0. 0.3 (0 (

SUMMARY OBJECTIVE: A survey among medical students in a Brazilian public university was performed to investigate the acceptance of organ donation in Brazil, particularly donation after circulatory death (DCD). METHODS: A questionnaire including 26 objectives and Likert scale questions was validated and sent to all medical students of our institution. The answers were analyzed considering the whole set of individuals as well as by dividing the medical students into two groups: less graduated students and more graduated students. RESULTS: From 1050 students, 103 spontaneous answers (9.8%) were retrieved after 3 weeks. A total of 89.3% agreed totally with deceased donor organ donation and 8.7% agreed partially. However, only 50.5% of the students agreed totally and 31.1% agreed partially to living donation. Students revealed that 82.6% know the concept of brain death. On the other hand, 71.8% of them declared not knowing the concept of planned withdrawal of life-sustaining therapy, mainly cardiorespiratory support. A total of 85.4% of students agreed totally with donation after brain death and 11.7% agreed partially. However, when questioned about donation in awaiting circulatory death after a planned withdrawal of life-sustaining therapy, only 18.4% agreed totally and 32% agreed partially. Both groups of less and more graduated students showed similar results. CONCLUSIONS: Our study found a clear lack of information and consequently in acceptance of DCD. Education in the field of end-of-life management may improve not only the acceptance of DCD donation but also the whole understanding of planned withdrawal of life-sustaining therapy.

RESUMO Objetivo Determinar a presença de glicosaminoglicanos na matriz extracelular do tecido conjuntivo colorretal neoplásico e não neoplásico, tendo em vista seu papel central no desenvolvimento e na progressão dos tumores. Métodos Amostras de tecidos colorretais neoplásicos e não neoplásicos foram obtidas de 64 pacientes operados com carcinoma colorretal sem metástases a distância. As expressões de heparan sulfato, sulfato de condroitina e sulfato de dermatan e seus fragmentos foram analisadas por espectrometria de massa por ionização por electrospray, com técnica de extração e quantificação de glicosaminoglicanos após proteólise e eletroforese. Para análise estatística, utilizaram-se média, desvio padrão e teste t de Student. Resultados Em gel de agarose, os glicosaminoglicanos extraídos de tecido colorretal mostraram três bandas eletroforéticas. A espectrometria de massa por ionização por electrospray mostrou fragmentos de dissacarídeos característicos de glicosaminoglicanos e indicou sua característica estrutural. Alguns picos na espectrometria de massa por ionização por electrospray não foram caracterizados como fragmentos de açúcares, sugerindo a presença de fragmentos de proteínas estruturais dos proteoglicanos, formadas durante a purificação dos glicosaminoglicanos. A quantidade média de condroitina e dermatan aumentou no tecido neoplástico em relação ao tecido normal (p=0,01). Por outro lado, a quantidade média de heparan foi menor no tecido neoplásico em relação ao tecido normal (p=0,03). Conclusão O método empregado permitiu determinar o perfil estrutural dos glicosaminoglicanos nas amostras. Tecidos neoplásicos apresentaram maiores quantidades de sulfato de condroitina e sulfato de dermatan em comparação com os não neoplásicos, enquanto o sulfato de heparan foi encontrado em menores quantidades nos tecidos neoplásicos.

ABSTRACT Objective To determine the presence of glycosaminoglycans in the extracellular matrix of connective tissue from neoplastic and non-neoplastic colorectal tissues, since it has a central role in tumor development and progression. Methods Tissue samples from neoplastic and non-neoplastic colorectal tissues were obtained from 64 operated patients who had colorectal carcinoma with no distant metastases. Expressions of heparan sulphate, chondroitin sulphate, dermatan sulphate and their fragments were analyzed by electrospray ionization mass spectrometry, with the technique for extraction and quantification of glycosaminoglycans after proteolysis and electrophoresis. The statistical analysis included mean, standard deviation, and Student’st test. Results The glycosaminoglycans extracted from colorectal tissue showed three electrophoretic bands in agarose gel. Electrospray ionization mass spectrometry showed characteristic disaccharide fragments from glycosaminoglycans, indicating their structural characterization in the tissues analyzed. Some peaks in the electrospray ionization mass spectrometry were not characterized as fragments of sugars, indicating the presence of fragments of the protein structure of proteoglycans generated during the glycosaminoglycan purification. The average amount of chondroitin and dermatan increased in the neoplastic tissue compared to normal tissue (p=0.01). On the other hand, the average amount of heparan decreased in the neoplastic tissue compared to normal tissue (p= 0.03). Conclusion The method allowed the determination of the glycosaminoglycans structural profile in colorectal tissue from neoplastic and non-neoplastic colorectal tissue. Neoplastic tissues showed greater amounts of chondroitin sulphate and dermatan sulphate compared to non-neoplastic tissues, while heparan sulphate was decreased in neoplastic tissues.

CONTEXTO E OBJETIVO: A heparanase-1 degrada heparam sulfato e está relacionada à progressão de tumor. Apesar de a isoforma heparanase-2 não possuir atividade catalítica, parece ser importante para modular a atividade da heparanase-1. A catepsina B é uma proteinase envolvida na metástase de tumores. O objetivo deste estudo foi analisar a expressão das isoformas de heparanase e atividade da catepsina B em amostras de plasma de pacientes com carcinomas gastrointestinais, comparando-se com indivíduos saudáveis (grupo controle). TIPO DE ESTUDO E LOCAL: Este é um estudo transversal analítico. Foram coletadas amostras de sangue periférico, em hospital público brasileiro, de 21 pacientes com diagnóstico histopatológico de carcinoma gastrointestinal e 43 indivíduos saudáveis. As análises foram realizadas em duas faculdades de medicina brasileiras. MÉTODOS: As isoformas da heparanase foram identificadas e quantificadas em amostras de plasma por Western blot. As atividades enzimáticas de heparanase-1 e catepsina B foram também mensuradas. RESULTADOS: Os resultados demonstraram que as expressões das isoformas de heparanase foram significativamente maiores nas amostras de plasma de pacientes com carcinoma gastrointestinal em comparação com o grupo controle. A análise feita por regressão logística mostra que aumento na expressão de heparanase-1 e heparanase-2 é exclusivamente dependente da presença de tumor. Houve aumento significativo na atividade da heparanase-1 e catepsina B no plasma dos pacientes. CONCLUSÃO: A superexpressão de heparanase-1 e heparanase-2, bem como o aumento da atividade de heparanase-1 e catepsina B no plasma, está associada com o diagnóstico de carcinoma gastrointestinal. Tais achados dão suporte ao uso de ensaios não invasivos (amostras de plasma) como método auxiliar para o diagnóstico de tumores gastrointestinais.

CONTEXT AND OBJECTIVE: Heparanase-1 degrades heparan sulfate and has been correlated with tumor progression. Although the isoform heparanase-2 has no catalytic activity, it seems to be important for modulating heparanase-1 activity. Cathepsin B is a proteinase involved in tumor metastasis. The aim of this study was to analyze heparanase isoform expression and cathepsin B activity in plasma samples from patients with gastrointestinal carcinomas, compared with healthy individuals (control group). DESIGN AND SETTING: This was an analytical cross-sectional study. Peripheral blood samples were collected at a Brazilian public hospital, from 21 patients with histopathological diagnoses of gastrointestinal carcinomas and from 43 healthy individuals. The analyses were performed in two Brazilian medical schools. METHODS: Heparanase isoforms were identified and quantified in plasma samples by means of Western blot. The enzymatic activities of heparanase-1 and cathepsin B were also measured. RESULTS: The results demonstrated that the expression of both heparanase isoforms was significantly greater in plasma samples from gastrointestinal carcinoma patients, compared with the control group. Logistic regression analysis showed that increased heparanase-1 and heparanase-2 expression was exclusively dependent on the tumor. There was a significant increase in heparanase-1 and cathepsin B activity in the patients' plasma. CONCLUSION: Overexpression of heparanase-1 and heparanase-2, along with increased heparanase-1 and cathepsin B activity in plasma, is associated with the diagnosis of gastrointestinal carcinoma. These findings provide support for using non-invasive assays (plasma samples) as an auxiliary method for diagnosing gastrointestinal tumors.

OBJECTIVE: This study sought to identify the relationship between fibroblast telomerase expression, myofibroblasts, and telomerase-mediated regulatory signals in idiopathic pulmonary fibrosis. METHODS: Thirty-four surgical lung biopsies, which had been obtained from patients with idiopathic pulmonary fibrosis and histologically classified as usual interstitial pneumonia, were examined. Immunohistochemistry was used to evaluate fibroblast telomerase expression, myofibroblast α-smooth muscle actin expression and the tissue expression of inter leu kin-4, transforming growth factor-β, and basic fibroblast growth factor. The point-counting technique was used to quantify the expression of these markers in unaffected, collapsed, mural fibrosis, and honeycombing areas. The results were correlated to patient survival. RESULTS: Fibroblast telomerase expression and basic fibroblast growth factor tissue expression were higher in collapsed areas, whereas myofibroblast expression and interleukine-4 tissue expression were higher in areas of mural fibrosis. Transforming growth factor-β expression was higher in collapsed, mural fibrosis and honeycombing areas in comparison to unaffected areas. Positive correlations were found between basic fibroblast growth factor tissue expression and fibroblast telomerase expression and between interleukin-4 tissue expression and myofibroblast α-smooth muscle actin expression. Negative correlations were observed between interleukin-4 expression and basic fibroblast growth factor tissue expression in areas of mural fibrosis. Myofibroblast α-smooth muscle actin expression and interleukin-4 tissue expression in areas of mural fibrosis were negatively associated with patient survival. CONCLUSION: Fibroblast telomerase expression is higher in areas of early remodeling in lung tissues demonstrating typical interstitial pneumonia, whereas myofibroblast α-smooth muscle actin expression predominates in areas of late remodeling. These events seem to be regulated by basic fibroblast growth factor and interleukin-4 tissue expression, respectively.

OBJECTIVE: We aimed to evaluate whether the inclusion of videothoracoscopy in a pleural empyema treatment algorithm would change the clinical outcome of such patients. METHODS: This study performed quality-improvement research. We conducted a retrospective review of patients who underwent pleural decortication for pleural empyema at our institution from 2002 to 2008. With the old algorithm (January 2002 to September 2005), open decortication was the procedure of choice, and videothoracoscopy was only performed in certain sporadic mid-stage cases. With the new algorithm (October 2005 to December 2008), videothoracoscopy became the first-line treatment option, whereas open decortication was only performed in patients with a thick pleural peel (>2 cm) observed by chest scan. The patients were divided into an old algorithm (n = 93) and new algorithm (n = 113) group and compared. The main outcome variables assessed included treatment failure (pleural space reintervention or death up to 60 days after medical discharge) and the occurrence of complications. RESULTS: Videothoracoscopy and open decortication were performed in 13 and 80 patients from the old algorithm group and in 81 and 32 patients from the new algorithm group, respectively (p<0.01). The patients in the new algorithm group were older (41 +1 vs. 46.3+ 16.7 years, p = 0.014) and had higher Charlson Comorbidity Index scores [0(0-3) vs. 2(0-4), p = 0.032]. The occurrence of treatment failure was similar in both groups (19.35% vs. 24.77%, p = 0.35), although the complication rate was lower in the new algorithm group (48.3% vs. 33.6%, p = 0.04). CONCLUSIONS: The wider use of videothoracoscopy in pleural empyema treatment was associated with fewer complications and unaltered rates of mortality and reoperation even though more severely ill patients were subjected to videothoracoscopic surgery.

OBJETIVO: Investigar a evolução clínica do transplante de intestino delgado ortotópico em ratos não-isogênicos. MÉTODOS: Setenta e dois ratos Wistar não-isogênicos, com peso variando entre 250 e 300g, foram utilizados como doadores e receptores em 36 transplantes ortotópicos de intestino delgado sem regime de imunossupressão. Os enxertos foram implantados nos receptores por meio de anastomose microvascular término-lateral aorta-aorta e porto-cava. A duração do procedimento, evolução clínica dos animais e sobrevida foram avaliados. Sobrevida menor que quatro dias foi considerada falha técnica. Os receptores foram sacrificados quando apresentaram sinais de rejeição grave do enxerto ou sobrevida maior que 120 dias. Necropsias foram realizadas em todos os receptores para avaliar alterações histopatológicas no enxerto. RESULTADOS: O tempo médio para o procedimento foi de 107 minutos. Seis receptores (16,7%) apresentaram falha técnica Vinte e sete receptores (75%) foram sacrificados por rejeição sendo dezenove (52,7%) entre o 7º e 15º dia de pós-operatório e oito (22,2%) entre o 34º e 47º. Análise histopatológica confirmou rejeição celular aguda severa nesses recipientes. Evolução sem complicações e sobrevida maior que 120 dias sem sinais de rejeição foi observada em três receptores (8,3%). CONCLUSÃO: O transplante de intestino delgado ortotópico em ratos Wistar não-isogênicos sem regime de imunossupressão apresenta evolução clínica variada.

PURPOSE: To investigate the clinical evolution of orthotopic small bowel transplantation in outbred rats. METHODS: Seventy-two outbred Wistar rats weighting from 250 to 300g were used as donor and recipient in 36 consecutives ortothopic small intestine transplantation without immunosuppression. The graft was transplanted into the recipient using end-to-side aortic and portacaval microvascular anastomosis. Procedure duration, animal clinical course and survival were evaluated. Survival shorter than four days was considered technical failure. Recipients were sacrificed with signs of severe graft rejection or survival longer than 120 days. Necropsies were performed in all recipients to access histopathological changes in the graft. RESULTS: Median time for the procedure was 107 minutes. Six recipients (16.7%) presented technical failure. Twenty-seven recipients were sacrificed due to rejection, being nineteen (52.7%) between 7th and 15th postoperative day and eight (22.2%) between 34th and 47th postoperative day. Graft histology confirmed severe acute cellular rejection in those recipients. Uneventful evolution and survival longer than 120 days without rejection were observed in three recipients (8.3%). CONCLUSION: Intestinal transplantation in outbred rats without immunosuppressant regiment accomplishes variable clinical evolution.

CONTEXTO: A recidiva de varizes em membros inferiores é complicação frequente da safenectomia e sua incidência atinge até 80% dos casos. OBJETIVO: Avaliar a sensibilidade do exame físico e do mapeamento com eco-color Doppler no diagnóstico da insuficiência do coto da veia safena magna, em doentes previamente operados, comparando-os com os achados da exploração operatória da junção safeno-femoral. MÉTODOS: Foram estudados prospectivamente 30 doentes envolvendo 37 membros submetidos previamente à safenectomia magna para tratamento de varizes e que foram reoperados por recidiva de varizes na região inguinal ou em face anterossuperior da coxa. Todos os doentes foram submetidos ao mapeamento com eco-color Doppler. Os dados foram comparados com os achados da exploração da crossa da veia safena magna na reoperação. RESULTADOS: A sensibilidade do mapeamento com eco-color Doppler para a presença de insuficiência do coto da veia safena magna foi de 70,3% (26 concordâncias dentre os 37 membros) e resultados falsos negativos ocorreram em 29,7% (11) membros avaliados (p=0,03). A sensibilidade do achado de varizes na região inguinal e na face anteromedial da coxa com a presença de insuficiência do coto da veia safena magna foi de 100% (37 concordâncias dentre os 37 membros) e não houve resultados falsos negativos. CONCLUSÕES: No doente já submetido à safenectomia magna, a presença no exame físico de varizes recidivadas em região inguinal e na face anteromedial da coxa é sugestivo de insuficiência do coto da veia safena magna, devendo-se realizar o mapeamento com eco color Doppler para o adequado planejamento da reexploração da crossa da veia safena magna.

BACKGROUND: Recurrent varicose veins of the lower limbs are a frequent complication of saphenous vein stripping and their incidence may reach 80% of the cases. OBJECTIVE: To evaluate the sensitivity of physical examination and color-Doppler flow mapping for the diagnosis of insufficiency in the greater saphenous vein stump in patients who had previously undergone saphenous vein stripping, comparing the results with the findings at reoperation of the saphenofemoral junction. METHODS: This prospective study included 37 limbs of 30 patients who had been previously submitted to saphenous vein stripping to treat varicose veins. Color-Doppler flow mapping was performed in all patients. Results were compared with findings at reoperation of the saphenofemoral junction. RESULTS: The sensitivity of color-Doppler flow mapping was 70.3% (26 limbs out of 37 limbs) and false negative results occurred in 11 (29.7%) limbs (p=0.03). The sensitivity of finding varicose veins in the groin and/or in the anteromedial aspect of the upper thigh was 100% (37 limbs out of 37 limbs), without false-negatives. CONCLUSIONS: In patients previously submitted to saphenous vein stripping, recurrent varicose veins found at physical examination in the groin or anteromedial aspect of the upper thigh are suggestive of greater saphenous vein stump insufficiency. Color-Doppler flow mapping should be performed for an adequate saphenofemoral junction reoperation.

OBJETIVO: Apresentar modelo de equipamento para anestesia inalatória em ratos, com melhor controle do fluxo, das perdas de anestésico na indução, da manutenção e da recuperação anestésica. MÉTODOS: O equipamento é constituído por compressor de ar com saída dupla, câmara de vidro fechada para indução, reservatório de vidro para o agente anestésico, máscara de inalação pediátrica, torneira de três vias, conexão em "Y" e sondas uretrais. Trezentos ratos Wistar (Rattus norvegicus albinus) foram anestesiados por via inalatória. Os parâmetros avaliados foram: viabilidade do equipamento, tempo de cada fase, reflexo corneano, tônus muscular e movimentos respiratórios na indução e na manutenção anestésica, além do volume de anestésico. RESULTADOS: A média do tempo gasto na indução anestésica foi de 7,3 minutos. A média do tempo de recuperação anestésica foi de 6,4 minutos. A média do volume de éter utilizado foi de 6,5 ml/h. Não foram observadas neste experimento morte dos animais, aumento excessivo de secreção traqueobrônquica ou interrupção da operação por superficialização anestésica. CONCLUSÃO: O equipamento proposto é prático, possui baixo custo e permite o controle adequado dos parâmetros de controle da anestesia durante todo o procedimento, tornando a anestesia inalatória em ratos segura e praticamente isenta de complicações.

PURPOSE: To introduce a model of equipment for inhalation anesthesia in rats that offers better control of both flow and losses of ether during induction, maintenance, and recuperation. METHODS: The equipment consists of an air compressor with two outlets, a closed glass induction chamber, a glass reservoir for the anesthetic agent, a pediatric inhalation mask, a three-way stopcock, a Y-connector, and urinary catheters. Three hundred Wistar rats (Rattus norvegicus albinus) were given inhalation anesthesia. The evaluated parameters were equipment operation, duration of each phase of anesthesia, corneal reflex, muscular tonus, respiration during induction and maintenance, and volume of anesthesia. RESULTS: The average time taken for induction was 7.3 minutes; the average anesthetic recuperation time was 6.4 minutes. The amount of anesthetic used varied according to the weight of the animal, with the average volume of ether used being 6.5ml/hour. The availability of oxygen (room air) decreased the recuperation time and averted both respiratory depression and insufficient depth of anesthesia. CONCLUSION: The proposed equipment is practical, inexpensive, and allows for satisfactory control of anesthetic parameters during the entire procedure, making inhalation anesthesia in rats safe and essentially complication free.