© F e r r a t a S t o r t i F o u n d a t i o npermeabilized in ice-cold PERM buffer III, washed in staining buffer and stained with the following antibodies: CD79a-APC; Akt-Alexa Fluor 488, PTEN-PE, phospho-Akt (S473)-Alexa Fluor 488, phospho-Akt (T308)-PE, and phospho-STAT5 (Y694)-Alexa Fluor 488. Samples were analyzed on a FACSAria or LSRFortessa using the gating strategy indicated in the Online Supplementary Appendix (Online Supplementary Figure S1). Endogenous PTEN in vitro lipid phosphatase assayPTEN phosphatase activity was measured in vitro as previously described. 13Endogenous CK2 in vitro kinase assay CK2 activity was measured in vitro as previously described. 15 Treatment with signaling inhibitorsCells were cultured in control medium or in the presence of CX-4945 or LY294002 for the indicated time points and used for protein and viability analysis. ImmunoblottingCell lysates were resolved by SDS-PAGE, transferred onto nitrocellulose membranes, and immunoblotted with the antibodies against actin, phospho-PTEN (S380), PTEN, CK2α and CK2α'. Analysis of cell viability and apoptosisCell viability was determined by double-staining with APC or FITC-conjugated Annexin V and propidium iodide (PI) and flow cytometry analysis, as previously described. 16 Statistical analysisDifferences between populations were calculated using unpaired two-tailed Students's t-test or Mann-Whitney test, as appropriate. Correlations were analyzed using the Pearson correlation coefficient. P<0.05 was considered significant. Results JAK/STAT and PI3K/Akt pathways are hyperactivated in adult B-ALL cellsHyperactivation of signaling pathways involved in promotion of proliferation and survival is commonly associated with cancer progression. Previous studies have shown that one of these signaling cascades, the JAK/STAT pathway, is constitutively activated in B-ALL patients displaying the BCR-ABL fusion (also known as Philadelphia chromosome (Ph)-positive cases) or CRFL2 overexpression in combination or not with activating mutations in JAK1 and JAK2. 3,17 Using phospho-specific flow cytometry and a gating strategy that enabled us to focus on blast cells (Online Supplementary Figure S1) to compare primary bone marrow cells from healthy donors with B-ALL blasts collected from leukemia patients at diagnosis (Table 1), we con-PI3K/Akt pathway activation in adult ALL haematologica | 2014; 99(6) 1063 TdT+, cCD79a+,CD19+, TdT+,CD10+, cCD79a+,CD19+, TdT+,cyIgM+, © F e r r a t a S t o r t i F o u n d a t i o n firmed that adult leukemia cases displayed constitutive hyperactivation of the JAK/STAT pathway ( Figure 1A; Online Supplementary Figures S1 and S2). Moreover, we found a discrete subgroup of samples that presented very high levels of STAT5 phosphorylation. In line with the knowledge that BCR-ABL drives STAT5 activation, 17,18 this group was enriched in Ph-positive cases ( Figure 1A, red labels).In contrast to JAK/STAT pathway, the activation status of PI3K/Akt signaling pathway and its potential role in adult ALL remain less well...

Women with mutations in the breast cancer genes BRCA1 or BRCA2 have an increased lifetime risk of developing breast, ovarian and other BRCA-associated cancers. However, the number of detected germline mutations in families with hereditary breast and ovarian cancer (HBOC) syndrome is lower than expected based upon genetic linkage data. Undetected deleterious mutations in the BRCA genes in some high-risk families are due to the presence of intragenic rearrangements such as deletions, duplications or insertions that span whole exons. This article reviews the molecular aspects of BRCA1 and BRCA2 rearrangements and their frequency among different populations. An overview of the techniques used to screen for large rearrangements in BRCA1 and BRCA2 is also presented. The detection of rearrangements in BRCA genes, especially BRCA1, offers a promising outlook for mutation screening in clinical practice, particularly in HBOC families that test negative for a germline mutation assessed by traditional methods.

Azul de metileno no tratamento da síndrome vasoplégica em cirurgia cardíaca. Quinze anos de perguntas, respostas, dúvidas e certezasMethylene blue for vasoplegic syndrome treatment in heart surgery. Fifteen years of questions, answers, doubts and certainties AbstractObjective: There is strong evidence that methylene blue (MB), an inhibitor of guanylate cyclase, is an excellent therapeutic option for vasoplegic syndrome (VS) treatment in heart surgery. The aim of this article is to review the MB's therapeutic function in the vasoplegic syndrome treatment.Methods: Fifteen years of literature review.Results: 1) Heparin and ACE inhibitors are risk factors; 2) In the recommended doses it is safe (the lethal dose is 40 mg/ kg); 3) The use of MB does not cause endothelial dysfunction; 4) The MB effect appears in cases of nitric oxide (NO) upregulation; 5) MB is not a vasoconstrictor, by blocking of the GMPc system it releases the AMPc system, facilitating the norepinephrine vasoconstrictor effect; 6) The most used dosage is 2 mg/kg as IV bolus followed by the same continuous infusion because plasmatic concentrations strongly decays in the first 40 minutes; 7) There is a possible "window of opportunity" for the MB's effectiveness.Conclusions: Although there are no definitive multicentric studies, the MB used to treat heart surgery VS, at the present time, is the best, safest and cheapest option, being a Brazilian contribution for the heart surgery.

Submitral left ventricular aneurysm is a cardiac pathology widely recognized, but relatively unknown, occurred almost exclusively in African black patients. Although still this idea of racial prevalence exists, cases have been described in patients of all the races. Ten Brazilian cases were reported. One of them was presented inside an Italian paper that refers the surgical treatment of a Brazilian patient of black race. We reported one more submitral left ventricular aneurysm case in a brown female patient, with antecedents of peripheral thromboembolism initially not identified as consequence of the cardiac pathology