Functional differences between two morphologically distinct cell subpopulations within a human colorectal carcinoma cell line

The LISP-I human colorectal adenocarcinoma cell line was isolated from a hepatic metastasis at the Ludwig Institute, São Paulo, SP, Brazil. The objective of the present study was to isolate morphologically different subpopulations within the LISP-I cell line, and characterize some of their behavioral aspects such as adhesion to and migration towards extracellular matrix components, expression of intercellular adhesion molecules and tumorigenicity in vitro. Once isolated, the subpopulations were submitted to adhesion and migration assays on laminin and fibronectin (crucial proteins to invasion and metastasis), as well as to anchorage-independent growth. Two morphologically different subpopulations were isolated: LISP-A10 and LISP-E11. LISP-A10 presents a differentiated epithelial pattern, and LISP-E11 is fibroblastoid, suggesting a poorly differentiated pattern. LISP-A10 expressed the two intercellular adhesion molecules tested, carcinoembryonic antigen (CEA) and desmoglein, while LISP-E11 expressed only low amounts of CEA. On the other hand, adhesion to laminin and fibronectin as well as migration towards these extracellular matrix proteins were higher in LISP-E11, as expected from its poorly differentiated phenotype. Both subpopulations showed anchorage-independent growth on a semi-solid substrate. These results raise the possibility that the heterogeneity found in the LISP-I cell line, which might have contributed to its ability to metastasize, was due to at least two different subpopulations herein identified.Fundação Antônio PrudenteUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de Microbiologia, Imunologia e ParasitologiaUNIFESP, EPM, Depto. de Microbiologia, Imunologia e ParasitologiaSciEL

Laminin concentration in ascites of patients with hepatic cirrhosis and peritoneal carcinomatosis

Laminin levels in ascitic fluid have been proposed as a marker for neoplastic ascites. We compared the concentration of laminin in serum and in ascitic fluid from patients with hepatic cirrhosis and peritoneal carcinomatosis and assessed the diagnostic value of serum laminin levels in differentiating neoplastic from benign ascites. Laminin concentrations were determined by ELISA with antibodies against laminin extracted from the human placenta, in patients with ascites due to peritoneal carcinomatosis (N = 20) and hepatic cirrhosis (N = 33). Patients with infected or hemorrhagic ascites were excluded. The receiver operating characteristic curve was used to determine the sensitivity and specificity of serum laminin for the diagnosis of neoplastic ascites. When compared to the group with cirrhosis, the carcinomatosis group presented significantly higher mean laminin levels in serum (3.3 ± 0.5 vs 2.1 ± 0.4 µg/ml, mean ± SD, P 2.25 µg/ml showed 100% sensitivity and 73% specificity for the diagnosis of neoplastic ascites. Serum concentration seems to be the main determinant of laminin levels in ascitic fluid and its values can be used as a diagnostic parameter in the study of neoplastic ascites.Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Disciplina de GastroenterologiaUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Disciplina de ImunologiaUNIFESP, EPM, Disciplina de GastroenterologiaUNIFESP, EPM, Disciplina de ImunologiaSciEL

Clubes de convergência nos municipios mineiros: uma análise via modelo threshold

This paper is makes an analysis of the process of convergence of income between the municipalities of the Minas Gerais (Brazil) in the period of 1980 to 2005, trying to identify the existence of clubs of convergence using a model threshold. Results indicated for the formation of four clubs of convergence, among them shows a process of convergence between the all clubs. Human capital proved to be significant in explaining the process of growth of the intermediate groups of municipalities richer. The results also indicated that the physical capital was significantly negative for explaining the process of economic growth of most rich clubs of convergence, demonstrating in this way, a negative role for public investment undertaken in the districts of the region.Income Convergence, Threshold Effect, Minas Gerais (Brazil), Community/Rural/Urban Development, Research Methods/ Statistical Methods, R11,

A Large-Area RPC Detector for Muography

A muon telescope equipped with four Resistive Plate Chambers of 2 m$^{2}$ per plane was tested with the muon scattering tomography technique. The telescope was operated during several hours with high atomic number materials located at its center with two detector planes on each side. With an intrinsic efficiency above 98%, spatial resolution around 1 cm and detector planes spaced by 45 cm, it was possible to identify the presence of a 5 cm thick tungsten block in 10 minutes of acquisition. The results obtained after five hours of acquisition are also presented in this communication

Advances Towards a Large-Area, Ultra-Low-Gas-Consumption RPC Detector

Large Resistive Plate Chamber systems have their roots in High Energy Physics experiments at the European Organization for Nuclear Research: ATLAS, CMS and ALICE, where hundreds of square meters of both trigger and timing RPCs have been deployed. These devices operate with complex gas systems, equipped with re-circulation and purification units, which require a fresh gas supply of the order of 6 cm$^{3}$/min/m$^{2}$, creating logistical, technical and financial problems. In this communication, we present a new concept in the construction of RPCs which allowed us to operate a detector at ultra-low gas flow regime. With this new approach, the glass stack is encapsulated in a tight plastic box made of polypropylene, which presents excellent water vapor blocking properties as well as a good protection against atmospheric gases.Comment: submission of the proceedings for the 15th PISA conference PM202

Endothelial nitric oxide synthase uncoupling as a key mediator of melanocyte malignant transformation associated with sustained stress conditions

Melanoma cell lines and cells corresponding to premalignant melanocytes were established by our group after subjecting a nontumorigenic murine melanocyte lineage, melan-a, to sequential cycles of anchorage blockade. Previous results showed that in melan-a cells the superoxide level increases after such procedure. Superoxide production during melanocyte de-adhesion was inhibited by L-sepiapterin, the precursor of eNOS cofactor BH(4), and increased by the inhibitor of BH(4) synthesis, DAHP, hence indicating a partial uncoupling state of eNOS. the eNOS uncoupling seems to be maintained in cells derived from melan-a, because they present decreased nitric oxide and increased superoxide levels. the inhibition of superoxide production in Tm5 melanoma cells with L-sepiapterin reinforces their eNOS-uncoupled state. the maintenance of oxidative stress seems to be important in melanoma apoptosis resistance because Mn(III)TBAP, a superoxide scavenger, or L-sepiapterin renders Tm5 cells more sensitive to anoikis and chemotherapy. More importantly, eNOS uncoupling seems to play a pivotal role in melanocyte malignant transformation induced by sustained anchorage impediment, because no malignant transformation was observed when L-NAME-treated melanocytes were subjected to sequential cycles of de-adhesion. Our results show that uncoupled eNOS contributes to superoxide production during melanocyte anchorage impediment, contributing to anoikis resistance and malignant transformation. (C) 2011 Elsevier Inc. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Federal de São Paulo, Disciplina Imunol, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Disciplina Farmacol, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Disciplina Nefrol, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Disciplina Imunol, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Disciplina Farmacol, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Disciplina Nefrol, BR-04023900 São Paulo, BrazilFAPESP: 06/61293-1FAPESP: 05/60334-3FAPESP: 08/50366FAPESP: 09/03335-8Web of Scienc

Stenosis of reverse great saphenous vein graft in infrainguinal arterial revascularization

SummaryObjectiveThe aim of this study was to evaluate the prevalence of hemodynamically significant infrainguinal bypasses stenosis using reverse great saphenous vein graft.MethodsFrom March of 2008 to March of 2009, 56 infrainguinal bypasses were performed with reverse great saphenous vein graft in 56 patients. On the 30th post-operative day, 32 out of 56 patients were submitted to vascular ultrasonography. The prevalence of significant graft stenosis was determined. In addition, the diagnosis of stenosis was related to the clinical and surgical characteristics of the patients. The variables analyzed at the moment of diagnosis were the localization of the graft stenosis, the risk factors associated with stenosis and the association of vascular ultrasonography findings with ankle-brachial pressure index (ABI).ResultsThe overall prevalence of significant graft stenosis was 48.4%. Out of the total number of observed stenosis, 19.4% were considered severe, and 29% mild or moderate. There was no significant association between the presence of significant stenosis and the following variables: gender, diabetes, hypertension, smoking, hipercholesterolemia, graft diameter, site of the distal anastomosis, and graft composition. There was a weak agreement between ABI and vascular ultrasonography in detecting stenosis in general (K=0.30; CL95% 0.232 - 0.473; p=0.018). However, there was a substantial agreement in detecting severe stenosis (K=0.75; CL95% 0.655 - 0.811; p=0.0001).ConclusionThere was a high prevalence of stenosis on the 30th post-operative day, mostly localized in the proximal half of the vein graft. There was no significant association of stenosis with clinical and surgical factors analyzed. ABI and vascular ultrasonography had weak agreement with the diagnosis of stenosis in general and an important agreement for the diagnosis of severe stenosis