Perfil plasmático da ICAM-1 e da VCAM-1 no pós-operatório cardíaco de lactentes submetidos à circulação extracorpórea

Introdução – Grande parte da população de crianças portadoras de defeitos cardíacos congênitos torna-se criticamente doente durante o primeiro ano de vida e necessita tratamento cirúrgico. No entanto, durante cirurgias cardíacas, ocorre uma resposta inflamatória intensa desencadeada pelo período de circulação extracorpórea (CEC), o que provoca disfunção de órgãos e tecidos. Os neutrófilos assumem um papel importante e complexo neste período, envolvendo a ligação às células endoteliais, ativação e liberação de mediadores inflamatórios. Esse processo é iniciado e mantido através de moléculas de adesão específicas, como a molécula de adesão intercelular-1 (ICAM-1) e a molécula de adesão celular-vascular-1 (VCAM-1). Formas solúveis destas moléculas apresentam variações de seus níveis durante cirurgias cardíacas com uso de CEC. Há poucos dados na literatura relacionados ao comportamento destas moléculas após cirurgia cardíaca com uso de CEC em lactentes, estando seu significado no plasma ainda por ser decifrado. Objetivos – Mensurar os níveis plasmáticos das moléculas de adesão solúveis ICAM-1 e VCAM-1 em condições basais e após exposição ao circuito de CEC em lactentes submetidos à cirurgia cardíaca para correção de defeitos cardíacos congênitos. Comparar os níveis plasmáticos destas moléculas entre pacientes acianóticos e cianóticos. Métodos – Foram estudados 21 lactentes, durante o período de junho de 1998 a março de 1999, submetidos à cirurgia cardíaca com uso de CEC. Foram analisados os níveis plasmáticos da ICAM-1 e da VCAM-1 solúveis pelo método de ELISA, na indução anestésica, ao término da CEC e 8 e 26 horas após o término da CEC. Resultados – As patologias cardíacas congênitas mais comuns foram defeito do septo atrioventricular e tetralogia de Fallot. As médias de idade e de peso foram 6,6 meses e 5,8 Kg. As medianas dos tempos de CEC e de clampeamento de aorta foram, respectivamente, 87 e 53 minutos. Todos os lactentes utilizaram inotrópicos. As medianas dos tempos de intubação e de internação foram 72 horas e 21 dias. A taxa de mortalidade dos pacientes foi de 9,5%. Os níveis basais da ICAM-1 e da VCAM-1 foram significativamente mais elevados do que aqueles considerados normais (P<0,0001). Os níveis da ICAM-1 diminuíram significaticamente ao término da CEC (P<0,001), voltando a aumentar significativamente 8 horas após o término deste período (P<0,005), sem no entanto, alcançar os valores basais 26 horas depois. A VCAM-1 apresentou comportamento semelhante. No entanto, 26 horas após CEC houve diminuição significativa de seus níveis em relação ao valor identificado 8 horas após tal período (P<0,005). Não houve diferença estatisticamente significativa quanto aos valores das moléculas entre pacientes acianóticos e cianóticos (P>0,1). Não houve associação entre os valores das moléculas e as variáveis perioperatórias e os desfechos clínicos. Conclusões – Os níveis plasmáticos das moléculas de adesão solúveis ICAM-1 e VCAM-1 são aumentados em lactentes com cardiopatias congênitas acianóticas e cianóticas no seu estado basal. Os níveis plasmáticos de ICAM-1 e VCAM-1 solúveis variam após exposição ao circuito de CEC em lactentes submetidos à cirurgia cardíaca para correção de cardiopatias congênitas, apresentando um comportamento característico nestes pacientes. Não há diferenças no comportamento das moléculas entre pacientes acianóticos e cianóticos.

Aberrant expression of E-cadherin and beta-catenin proteins in placenta of bovine embryos derived from somatic cell nuclear transfer

Abnormal placental development is common in the bovine somatic cell nuclear transfer (SCNT)-derived fetus. In the present study, we characterised the expression of E-cadherin and beta-catenin, structural proteins of adherens junctions, in SCNT gestations as a model for impaired placentation. Cotyledonary tissues were separated from pregnant uteri of SCNT (n - 6) and control pregnancies (n - 8) obtained by artificial insemination. Samples were analysed by western blot, quantitative RT-PCR (qRT-PCR) and immunohistochemistry. Bovine trophectoderm cell lines derived from SCNT and control embryos were analysed to compare with the in utero condition. Although no differences in E-cadherin or beta-catenin mRNA abundance were observed in fetal tissues between the two groups, proteins encoded by these genes were markedly under-expressed in SCNT trophoblast cells. Immunohistochemistry revealed a different pattern of E-cadherin and total beta-catenin localisation in SCNT placentas compared with controls. No difference was observed in subcellular localisation of dephosphorylated active-beta-catenin protein in SCNT tissues compared with controls. However, qRT-PCR confirmed that the wingless (WNT)/beta-catenin signalling pathway target genes CCND1, CLDN1 and MSX1 were downregulated in SCNT placentas. No differences were detected between two groups of bovine trophectoderm cell lines. Our results suggest that impaired expression of E-cadherin and beta-catenin proteins, along with defective beta-catenin signalling during embryo attachment, specifically during placentation, is a molecular mechanism explaining insufficient placentation in the bovine SCNT-derived fetus.

Expressão da E-caderina em carcinoma de células escamosas e no tumor de células basais de cães

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expressão imuno-histoquímica das integrinas α2ß1, α3ß1, e α5ß1, em mucosa oral normal, hiperplasia fibroepitelial inflamatória oral e displasia epitelial oral

The objective of this study was perform by the streptoavidin-biotin technique an immunohistochemical analysis of α2β1, α3β1e α5β1 integrins in 11 normal oral mucosa (NOM), 16 oral inflammatory fibroepithelial hyperplasia (OIFH) and 25 oral epithelial dysplasia (OED) (16 mild, 2 moderates and 7 severe), to determine if exists qualitative alteration in the expression of these integrins and if this guard relation with the oral epithelial modifications. It was observed that for the α2β1 integrin the majority of the sample showed a predominantly intense labeling diffusely distributed in the intercellular contacts and the cytoplasm of cells of the basal and suprabasal layers, without difference of this profile between the different types of specimens, however with a trend to weak or loss of expression in 21.1% of the OEDs, being all the specimens that had not expressed this heterodimer, severe OEDs. For the α3β1 integrin the majority of the sample showed a weak or absent labeling in basal layer. The α5β1 integrin showed a predominant strong diffuse labeling in the intercellular contacts and cytoplasm in the suprabasal layer, with difference only in the labeling intensity between the types of specimens, inhabiting this difference in the OEDs, where 12 (48%) specimens had shown a weak labeling. It was concluded that the evaluated integrins can be involved in the cell-cell, cell-ECM interactions modulating the cellular differentiation and maintenance of the epithelial structural arrangement. The variable expression of the α5β1 integrin in the OEDs, could suggest, respectively, a role of this molecule in the cellular survival, with intention to perpetuate the modified phenotype in these lesions, or a suppressor role on the modified phenotype due to lack of interaction of this molecule with the fibronectina of the MEC

Expressão imuno-histoquímica da e-caderina e da ß-catenina em carcinoma epidermóide oral com e sem metástase nodal

The adhesion molecules E-cadherin and β-catenin have been studied as possible markers to distinguish carcinomas with and without metastatic potential. The objective of this research was to study the imunohistochemistry expression of the E-cadherin and β-catenin in oral squamous cell carcinoma (OSCC), aiming to contribute for the better understanding of the biological behavior of this lesion. The sample consisted of 30 cases of OSCC, being 15 of tongue and 15 of lower lip. The profile and intensity of labeling and semi quantitative analysis of the percentage of immunopositive tumoral cells in membrane for E-cadherin and β-catenin had been related with the anatomical localization of the lesion, the presence or not of nodal metastasis and the histological grade of malignancy in the invasive front area of the tumor. It was registered the presence or not of cytoplasmic and nuclear labeling of the β-catenin. The results had been submitted to the statistical analysis, being used the Mann-Whitney Test, the Fisher Test and the Spearman Correlation Coefficient (α=0, 05). The results showed that the expression in membrane for E-cadherin and β-catenin was, predominantly, the heterogeneous profile in the lower lip and tongue carcinomas, as well as in the cases with and without nodal metastasis. It was not observed significant statistical difference between expression profile and amount of immunopositive cells for E-cadherin, β-catenin and the anatomical localization of the lesion and for the presence or not of nodal metastasis. However, there was significant difference of the reduced expression of these proteins with the high score of malignancy. It was verified that the expression of the β-catenin in cytoplasm was present in 22 (73.33%) of the 30 analyzed cases, and 6 cases (20%) showed nuclear expression. The statistical analysis detected significant association between the expression of the β-catenin in the cytoplasm with the histological grade of malignancy, being this molecule more frequently present in the cytoplasm in the cases of high score of malignancy. It was concluded that the reduced immunoexpression of these proteins in membrane can be related with the lowest cellular differentiation, as well as with the pattern of invasion in nests and isolated cells, demonstrated in the cases of OSCC with high histological grade of malignancy

Determinação da expressão da molécula de adesão CD56 em plasmócitos no mieloma múltiplo através de estudo imuno-histoquímico

O papel das moléculas de adesão celular, na fisiopatologia do mieloma múltiplo (MM), tem sido alvo de vários estudos nos últimos anos. A expressão de CD56 pelos plasmócitos tumorais está associada a comportamento clínico menos agressivo da doença, e sua perda tem sido descrita na fase de leucemização plasmocitária. A determinação da expressão da molécula CD56 pelos plasmócitos tumorais pode ser obtida através de citometria em fluxo, revelando positividade em 55% a 78% dos casos. No presente estudo, objetivamos verificar a expressão da molécula CD56 por plasmócitos tumorais na medula óssea de portadores de MM, utilizando o estudo imuno-histoquímico das amostras histológicas obtidas ao diagnóstico. Analisamos as amostras de medula óssea de vinte portadores de MM, e realizamos o estudo imuno-histoquímico para determinação da expressão das cadeias leves kappa e lambda e da molécula CD56 pelos plasmócitos tumorais. A expressão da molécula CD56 foi importante em três casos, moderada em seis, discreta em quatro e negativa em sete. O estudo imuno-histoquímico mostrou-se válido para determinação da expressão de CD56 por plasmócitos tumorais em portadores de MM, fornecendo resultados semelhantes aos descritos para os obtidos por citometria em fluxo. Através do estudo imuno-histoquímico, foi possível observar variações da expressão da molécula CD56.

Atividade antiinflamatória de uma heterofucana da alga marrom Padina gymnospora

Fucans, sulfated polysaccharides extracted from brown algae and some echinoderms, have been extensively studied for its diverse biological activities and because of its interference with molecular mechanisms of cell to cell recognition, including leukocyte trafficking from blood vessels into sites of inflammation mediated by selectin, a family of adhesion molecules. In the present study, we examined structural features of a heterofucan extracted from brown algae Padina gymnospora and its effect on the leukocyte migration to the peritoneum. The sulfated polysaccharides were extracted from the brown seaweed by proteolysis with the proteolytic enzyme maxatase. The presence of protein and uronic acid contamination was detected in the crude polysaccharide extract. Fractionation of the crude extract with growing concentrations of acetone produced five fractions with different concentrations of fucose, xylose, uronic acid, galactose, glucose and sulfate. The fraction precipitated with 1.5 volumes of acetone was characterized by infrared and nuclear magnetic resonance, through which can be observed the presence of sulfate groups in the C4 of -L-fucose. The anti-inflammatory action of this composite was assessed by a sodium thioglycollate-induced peritonitis assay and through nitric oxide production by the peritoneal macrophages using Griess reagent. Fraction F1.5 was efficient in reducing leukocyte influx into the peritoneal cavity when 10 mg/kg and 25mg/kg were used, resulting in a decrease of 56 and 39%, respectively. A decrease of nitric oxide production occurred when high concentrations of fucana were used. The cytotoxicity of the composite was also assessed using the reduction of 3-(4,5 dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT). Fraction F1.5 had no cytotoxicity when 500 μg/mL of the fraction was used. This study suggests the use of fraction F1.5 (heterofucan) as an anti-inflammatory

Familial cancer: depressed NK-cell cytotoxicity in healthy and cancer affected members

Este estudo se reporta às funções de células natural killer (NK), como adesão, lise e citotoxicidade e de subpopulações de células T em uma família com alta prevalência de pacientes com câncer e que apresentaram: glioblastoma, leucemia mielóide crônica, osteoblastoma, melanoma e carcinomas gástrico, pancreático e cólon retal. Quinze membros dessa família foram estudados, sendo 13 sadios, acompanhados por 5 anos e dois com câncer: glioblastoma e leucemia mielóide crônica. Duas pessoas sadias, no momento da avaliação, desenvolveram posteriormente osteoblastoma mandibular ou melanoma maligno. Como controle, foram avaliados 19 indivíduos saudáveis de faixa etária equivalente. A determinação de linfócitos T CD3+ e de suas subpopulações CD4+ e CD8+ foi realizada empregando-se anticorpos monoclonais e a atividade citotóxica de células NK, avaliada pelo teste de single-cell contra células alvo da linhagem eritroleucêmica K562. Os resultados mostraram que as percentagens de células T totais (CD3+), da subpopulação CD4+ e da relação CD4/CD8 foram significativamente menores nos indivíduos da família estudada em comparação aos valores observados no grupo controle. em todos os membros dessa família a percentagem de formação de conjugados entre células NK-células alvo foi inferior ao valor mínimo observado nos controles. Essa alteração poderia estar relacionada a defeito na expressão de moléculas de adesão, presentes na membrana de células NK, como provável causa das alterações funcionais dessas células. A herança dos mecanismos determinantes desta deficiência pode ser um fator de risco, com valor prognóstico para o desenvolvimento de cancer.

Atherosclerosis morphology and pathogenesis

Atherosclerosis is a very common and important disease being the most important cause of mortality in Brazil. Indeed, in 1995, 23.3% of deaths, all ages, in our country, were the consequence of atherosclerosis. This percentage grows to 26.3% for S. Paulo and 32.7% for Rio Grande do Sul. Morphologically, there are 3 main types of lesions: fatty streaks, fibrous plaques, and complicated lesions. Fatty streaks are inocuous and occur early in life. In some persons, with age, they change into fibrous plaques that may lead to stenosis. They also may become complicated by erosion, calcification, hemorrhage and thrombosis. Atherosclerosis is initiated by endothelial functional alterations responsible for increase in permeability to macromolecules, adhesion, and migration of monocytes-macrophages and lymphocytes plus recruitment of platelets and smooth-muscle medial cells. Adhesion molecules, cytokines, growth factors, and free radicals are locally synthesized, favoring proliferation of extracellular matrix and progression of the lesion. Experimental, clinical, and epidemiological evidence point to the importance of lipids, mainly cholesterol-rich low-density lipoprotein (LDL), as one of the most important molecules involved in the genesis and progression of atherosclerosis. Patients with a genetic disorder of cholesterol metabolism (familial hyperlipidemia), caused by a decrease in the availability of receptors for LDL, develop severe atherosclerosis early in life. A series of other factors, such as age, diabetes melitus, diet, hypertension, lack of exercise, elevated hemocysteinemia, immunological disorders, and coagulation instability, are related to the progression of atherosclerosis. All of them are capable of altering the endothelium or increasing the offer of LDL. All the above-mentioned factors are systemic; but atherosclerosic lesions are focal, located at preferential sites such as the emergence of colaterals, bifurcations, and curvatures of arteries, all areas in which the laminar flow is disturbed. In these areas shear stress is diminished favoring the prolongation of permanence time of lipid particles, cells, cytokines, growth factors, etc., in the vicinity of the endothelium. Moreover, the endothelium has sensors that act as transducers of mechanical forces in biological responses. Experimental data demonstrate that the number and quality of adhesion molecules, cytokines, and growth factors synthetized, as well as the local production of radicals, and pro and anticoagulation factors may change with shear stress favoring or not the local establishment and progression of atherosclerotic lesions.

Myxopapillary ependymoma of the posterior mediastinum

A left paravertebral mass discovered incidentally on routine examination in a 39-year-old woman is described. Computerized tomography studies revealed a 7 × 6 cm, well circumscribed, noncalcified soft tissue mass with lobular borders abutting the left inferior pulmonary vein and descending aorta. It was not possible to determine the exact anatomic location of the mass based on the imaging studies as both peripheral lung tumors and posterior mediastinal lesions may exhibit the imaging findings described here. At thoracotomy, the mass was seen to be well circumscribed, focally attached to the pleura but without involvement of lung parenchyma, and situated in the left posterior mediastinum. On histological examination, the lesion showed the classical features of myxopapillary ependymoma. Immunohistochemical studies confirmed this impression by demonstrating strong positivity of the tumor cells for S-100 protein, glial fibrillary acidic protein, and CD99 and negative staining with other differentiation markers. A review of the literature with a discussion of the histologic and radiologic differential diagnosis of these lesions is presented. © 2006 Elsevier Inc. All rights reserved.

Gene expression changes associated with myocarditis and fibrosis in hearts of mice with chronic chagasic cardiomyopathy

Chronic chagasic cardiomyopathy is a leading cause of heart failure in Latin American countries. About 30% of Trypanosoma cruzi-infected individuals develop this severe symptomatic form of the disease, characterized by intense inflammatory response accompanied by fibrosis in the heart.We performed an extensive microarray analysis of hearts from a mouse model of this disease and identified significant alterations in expression of ~12% of the sampled genes. Extensive up-regulations were associated with immune-inflammatory responses (chemokines, adhesion molecules, cathepsins, and major histocompatibility complex molecules) and fibrosis (extracellular matrix components, lysyl oxidase, and tissue inhibitor of metalloproteinase 1). Our results indicate potentially relevant factors involved in the pathogenesis of the disease that may provide newtherapeutic targets in chronic Chagas disease. © 2010 by the Infectious Diseases Society of America.

Methylation pattern of THBS1, GATA-4, and HIC1 in pediatric and adult patients infected with helicobacter pylori

Background: Helicobacter pylori infection is usually acquired in childhood and persists into adulthood if untreated. The bacterium induces a chronic inflammatory response, which is associated with epigenetic alterations in oncogenes, tumor-suppressor genes, cell-cycle regulators, and cell-adhesion molecules. Aim: The aim of this study was to analyze the effect of H. pylori infection on the methylation status of Thrombospondin-1 (THBS1), Hypermethylated in cancer 1 (HIC1) and Gata binding protein-4 (GATA-4) in gastric biopsy samples from children and adults infected or uninfected with the bacterium and in samples obtained from gastric cancer patients. Methods: The methylation pattern was analyzed with methylation-specific PCR. Results: Our results showed that H. pylori infection was associated with methylation of the promoter regions of the THBS1 and GATA-4 genes in pediatric and adult samples (p < 0.01). HIC1 showed the lowest level of methylation, which was not an early event during gastric carcinogenesis. Conclusions: The results from this study indicate that methylation of THBS1 and GATA-4 occurs in the early stages of chronic gastritis and gastric cancer in association with H. pylori infection; however, in gastric cancer samples, other mechanisms cooperate with the down-regulation of these genes. Methylation of HIC1 may not be the principal mechanism implicated in its down-regulation in gastric cancer samples. © 2013 Springer Science+Business Media New York.

Expressão da e-caderina e do fator de crescimento do endotélio vascular no carcinoma de células escamosas e no tumor de células basais de cães

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo da resposta imune humoral em mulheres com neoplasia ovariana. -

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Padrão histológico, perfil imunoistoquímico e potencial pré-maligno das lesões diaplásicas da próstata canina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)