ANÁLISE DE DEMONSTRATIVOS FINANCEIROS DE MODO A IDENTIFICAR AS ALTERAÇÕES NA EVIDENCIAÇÃO DOS ATIVOS INTANGÍVEIS EM FUNÇÃO DA LEI Nº 11.638/07

With the internationalization of accounting standards, in Brazil was promulgated the law n° 11. 638/07 that created the intangible group in asset of balance sheet. This study has for objective to analyze alterations in evidencing form of intangible assets, based on financial statements of 10 companies with greater social capital disclosed by BM&FBOVESPA in2010, incomparison to evidenced elements before law 11.638/07. For development of study some methodological procedures were used, standing out the content analysis, which was accomplished in financial demonstratives: Balance Sheet, Notes to the Financial Statements and Management Report. As result was observed: (a) the "A" and "I" companies have the largest variety of intangible evidenced in demonstratives; (b) the intangible types more evidenced in period were "Franchise and licenses", Software" and "Agio for expectation about future profitability"; and (c) "Marks" were the intangible more evidenced implicitly in administration reports of entities in study.ive an)� �h���M�as theoretical and conceptual inductive logic. The study is also documentary and adopts the technique of content analysis. As main results: (i) in the national context, there is the empirical relative to theoretical, with more than 95.0% of the work from the perspective of Disclosure and 90.0% in the financial perspective are empirical (ii) identified that in the national strategic perspective with models for measuring and managing intellectual capital as well as multicriteria methodologies, while the international context, the Strategic perspective, there was the adoption of models based on resource and concept (RBV), knowledge-based (KBV), balance sheets or frameworks - Balanced Scorecard (BSC) and specific models of intellectual capital (IC), (iii) as to economic segments stand out in the national focus on education with 13 jobs (9.8%). In the international context, the segments: Financial, Banking, Industrial, and High Technology Companies were mostly received attention in relation to empirical studies, representing in this sense, more than 10.0% of the studies identified. We conclude that the theoretical discussions are relevant or related to the definitions suggested by Marr (2005) proposed the ten prospects in both contexts. Com a internacionalização das normas contábeis, no Brasil foi publicada a Lei n° 11.638/07, a qual criou o grupo intangível no ativo do balanço patrimonial. Este estudo tem por objetivo analisar as alterações em termos da evidenciação dos ativos intangíveis, nos demonstrativos contábeis das dez empresas com maior capital social divulgadas pela BM&FBOVESPA no ano de 2010, em comparação com os elementos evidenciados anterior a Lei nº 11.638/07. No estudo foram utilizados alguns procedimentos metodológicos como a análise de conteúdo realizada nos demonstrativos financeiros: Balanço Patrimonial, Notas Explicativas e Relatório da Administração. Como resultado, observou-se que: (a) as empresas “A” e “I” possuem a maior variedade de intangíveis evidenciados nos demonstrativos; (b) os tipos de intangíveis mais evidenciados no período foram “Franquias e licenças”, “Software” e “Ágio por expectativa de rentabilidade futura”; e (c) “Marcas” foi o intangível mais evidenciado implicitamente nos relatórios de administração das entidades em estudo.

Regulation of RasGRP via a Phorbol Ester-Responsive C1 Domain

As part of a cDNA library screen for clones that induce transformation of NIH 3T3 fibroblasts, we have isolated a cDNA encoding the murine homolog of the guanine nucleotide exchange factor RasGRP. A point mutation predicted to prevent interaction with Ras abolished the ability of murine RasGRP (mRasGRP) to transform fibroblasts and to activate mitogen-activated protein kinases (MAP kinases). MAP kinase activation via mRasGRP was enhanced by coexpression of H-, K-, and N-Ras and was partially suppressed by coexpression of dominant negative forms of H- and K-Ras. The C terminus of mRasGRP contains a pair of EF hands and a C1 domain which is very similar to the phorbol ester- and diacylglycerol-binding C1 domains of protein kinase Cs. The EF hands could be deleted without affecting the ability of mRasGRP to transform NIH 3T3 cells. In contrast, deletion of the C1 domain or an adjacent cluster of basic amino acids eliminated the transforming activity of mRasGRP. Transformation and MAP kinase activation via mRasGRP were restored if the deleted C1 domain was replaced either by a membrane-localizing prenylation signal or by a diacylglycerol- and phorbol ester-binding C1 domain of protein kinase C. The transforming activity of mRasGRP could be regulated by phorbol ester when serum concentrations were low, and this effect of phorbol ester was dependent on the C1 domain of mRasGRP. The C1 domain could also confer phorbol myristate acetate-regulated transforming activity on a prenylation-defective mutant of K-Ras. The C1 domain mediated the translocation of mRasGRP to cell membranes in response to either phorbol ester or serum stimulation. These results suggest that the primary mechanism of activation of mRasGRP in fibroblasts is through its recruitment to diacylglycerol-enriched membranes. mRasGRP is expressed in lymphoid tissues and the brain, as well as in some lymphoid cell lines. In these cells, RasGRP has the potential to serve as a direct link between receptors which stimulate diacylglycerol-generating phospholipase Cs and the activation of Ras

Irs2 Silencing Increases Apoptosis And Potentiates The Effects Of Ruxolitinib In Jak2v617f-positive Myeloproliferative Neoplasms.

The recurrent V617F mutation in JAK2 (JAK2V617F) has emerged as the primary contributor to the pathogenesis of myeloproliferative neoplasms (MPN). However, the lack of complete response in most patients treated with the JAK1/2 inhibitor, ruxolitinib, indicates the need for identifying pathways that cooperate with JAK2. Activated JAK2 was found to be associated with the insulin receptor substrate 2 (IRS2) in non-hematological cells. We identified JAK2/IRS2 binding in JAK2V617F HEL cells, but not in the JAK2WT U937 cell line. In HEL cells, IRS2 silencing decreased STAT5 phosphorylation, reduced cell viability and increased apoptosis; these effects were enhanced when IRS2 silencing was combined with ruxolitinib. In U937 cells, IRS2 silencing neither reduced cell viability nor induced apoptosis. IRS1/2 pharmacological inhibition in primary MPN samples reduced cell viability in JAK2V617F-positive but not JAK2WT specimens; combination with ruxolitinib had additive effects. IRS2 expression was significantly higher in CD34+ cells from essential thrombocythemia patients compared to healthy donors, and in JAK2V617F MPN patients when compared to JAK2WT. Our data indicate that IRS2 is a binding partner of JAK2V617F in MPN. IRS2 contributes to increased cell viability and reduced apoptosis in JAK2-mutated cells. Combined pharmacological inhibition of IRS2 and JAK2 may have a potential clinical application in MPN.

Bayesian multi-source regression and monocyte-associated gene expression predict BCL-2 inhibitor resistance in acute myeloid leukemia

The FDA recently approved eight targeted therapies for acute myeloid leukemia (AML), including the BCL-2 inhibitor venetoclax. Maximizing efficacy of these treatments requires refining patient selection. To this end, we analyzed two recent AML studies profiling the gene expression and ex vivo drug response of primary patient samples. We find that ex vivo samples often exhibit a general sensitivity to (any) drug exposure, independent of drug target. We observe that this "general response across drugs" (GRD) is associated with FLT3-ITD mutations, clinical response to standard induction chemotherapy, and overall survival. Further, incorporating GRD into expression-based regression models trained on one of the studies improved their performance in predicting ex vivo response in the second study, thus signifying its relevance to precision oncology efforts. We find that venetoclax response is independent of GRD but instead show that it is linked to expression of monocyte-associated genes by developing and applying a multi-source Bayesian regression approach. The method shares information across studies to robustly identify biomarkers of drug response and is broadly applicable in integrative analyses

The PI3K/Akt1 pathway enhances steady-state levels of FANCL

Fanconi anemia hematopoietic stem cells display poor self-renewal capacity when subjected to a variety of cellular stress. This phenotype raises the question of whether the Fanconi anemia proteins are stabilized or recruited as part of a stress response and protect against stem cell loss. Here we provide evidence that FANCL, the E3 ubiquitin ligase of the Fanconi anemia pathway, is constitutively targeted for degradation by the proteasome. We confirm biochemically that FANCL is polyubiquitinated with Lys-48-linked chains. Evaluation of a series of N-terminal-deletion mutants showed that FANCL's E2-like fold may direct ubiquitination. In addition, our studies showed that FANCL is stabilized in a complex with axin1 when glycogen synthase kinase-3β is overexpressed. This result leads us to investigate the potential regulation of FANCL by upstream signaling pathways known to regulate glycogen synthase kinase-3β. We report that constitutively active, myristoylated-Akt increases FANCL protein level by reducing polyubiquitination of FANCL. Two-dimensional PAGE analysis shows that acidic forms of FANCL, some of which are phospho-FANCL, are not subject to polyubiquitination. These results indicate that a signal transduction pathway involved in self-renewal and survival of hematopoietic stem cells also functions to stabilize FANCL and suggests that FANCL participates directly in support of stem cell function

Omecamtiv mecarbil in chronic heart failure with reduced ejection fraction, GALACTIC‐HF: baseline characteristics and comparison with contemporary clinical trials

Aims: The safety and efficacy of the novel selective cardiac myosin activator, omecamtiv mecarbil, in patients with heart failure with reduced ejection fraction (HFrEF) is tested in the Global Approach to Lowering Adverse Cardiac outcomes Through Improving Contractility in Heart Failure (GALACTIC‐HF) trial. Here we describe the baseline characteristics of participants in GALACTIC‐HF and how these compare with other contemporary trials. Methods and Results: Adults with established HFrEF, New York Heart Association functional class (NYHA) ≥ II, EF ≤35%, elevated natriuretic peptides and either current hospitalization for HF or history of hospitalization/ emergency department visit for HF within a year were randomized to either placebo or omecamtiv mecarbil (pharmacokinetic‐guided dosing: 25, 37.5 or 50 mg bid). 8256 patients [male (79%), non‐white (22%), mean age 65 years] were enrolled with a mean EF 27%, ischemic etiology in 54%, NYHA II 53% and III/IV 47%, and median NT‐proBNP 1971 pg/mL. HF therapies at baseline were among the most effectively employed in contemporary HF trials. GALACTIC‐HF randomized patients representative of recent HF registries and trials with substantial numbers of patients also having characteristics understudied in previous trials including more from North America (n = 1386), enrolled as inpatients (n = 2084), systolic blood pressure < 100 mmHg (n = 1127), estimated glomerular filtration rate < 30 mL/min/1.73 m2 (n = 528), and treated with sacubitril‐valsartan at baseline (n = 1594). Conclusions: GALACTIC‐HF enrolled a well‐treated, high‐risk population from both inpatient and outpatient settings, which will provide a definitive evaluation of the efficacy and safety of this novel therapy, as well as informing its potential future implementation

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Expression cloning of RasGRP : a member of the CDC25 family of Ras exchangers

The fact that Ras mutations are found in approximately 30% of all human malignancies points towards a role for Ras signalling pathways in the process of malignant transformation. An expression cloning strategy was used to identify genes that were equal to Ras in their ability to transform cells. Upon screening a T hybridoma cell cDNA library we identified a novel cDNA whose overexpression in NIH 3T3 cells caused transformation phenotypically similar to that of Ras overexpression. RasGRP, the protein encoded by this cDNA, contained several unique structural features, the foremost being regions of high sequence similarity to the CDC25 family of guanine nucleotide exchange factors (GNEFs). RasGRP not only contained the three structurally conserved regions found in these domains but also contained a Ras exchange motif (REM) box that is found in GNEFs that are capable of exchange on true Ras proteins. Overexpression of RasGRP caused hyperactivation of MAP kinases, specifically ERK 1 and 2. This data is compatible with the idea RasGRP is capable of causing Ras activation. Furthermore, the ability of RasGRP to transform cells and activate MAP kinase was eliminated by mutations affecting the exchange factor domain or by deletions of the REM box. RasGRP also contained a Cl domain which was very similar in sequence to diacylglycerol- and phorbol ester-binding Cl domains of protein kinase Cs (PKCs). RasGRP's Cl domain was found to be essential for fibroblast transformation and could be functionally replaced by a prenylation signal or by the Cl domain of PKCδ. RasGRP was found to localize to the ER as well as other membranes and this localization was dependent upon the Cl domain. The isolated Cl domain was also capable of re-localization in response to phorbol ester stimulation and to increases in diacylglycerol produced upon phosphatidylcholine-phospholipase C stimulation. These responses were identical to those of the PKC5-C1 domain. Taken altogether, these results provide the evidence on which to base the following model of Ras activation via RasGRP. It states that RasGRP's Cl domain facilitates the translocation of RasGRP to membranes enriched in diacylglycerol or phorbol ester. Once at the membrane, RasGRP is able to interact with Ras proteins tethered there by prenylation. GDP exchange then takes place via RasGRP's REM/GEF domains and the Ras pathway is activated. RasGRP also contains a pair of calcium binding EF hands, a proline rich region, and an a-helical domain. These domains were found to be non-essential for fibroblast transformation and it will be of interest to discover what roles they play in RasGRP function. RasGRP exhibits a unique pattern of expression when compared to other GNEFs capable of exchange on Ras. It is expressed extensively in brain, thymus, spleen, and bone marrow, which points to possible roles in neuronal and lymphoid signalling.Medicine, Faculty ofMedical Genetics, Department ofGraduat

BREVETTO/PATENT FI2005A000038. TITOLO: OSTEOBLASTI UMANI INGEGNERIZZATI CHE ESPRIMONO LA PROTEINA FLUORESCENTE eGFP

none4Le cellule umane ingegnerizzate sono osteoblasti che esprimono in maniera costitutiva la proteina verde fluorescente. Questa linea cellulare può essere impiegata per determinare la biocompatibilità di materiali come ad esempio protesi in ortopedia e odontoiatria.Depositato il 11 Marzo 2005noneMORELLI C; CAMPIONI K; CIANNILLI A; TOGNON M.Morelli, Cristina; Campioni, Katia; Ciannilli, Alessandra; Tognon, Maur

BREVETTO INTERNAZIONALE. INTERNATIONAL PATENT PCT/EP2006/060603. Engineered human osteoblast cells expressing the fluorescent eGFP

Title A new genetically engineered human cell line for the in vitro characterization of biomaterials. Description In the dentistry and orthopaedic perspectives tissue engineering is focused on the development of innovative materials whose action consists in recruiting bone progenitor cells and in stimulating their proliferation. Doing so, they favourite the development of the tissue which should be replaced. In this context, it is clear that these materials should not only allow cells adhesion and proliferation, but also ensure that the attached cells will maintain the histological features of the original tissue. Careful attention should therefore be addressed to characterize the influence of the biomaterial on bone cells behaviour and, in turn, on bone tissue formation. The ultimate approach to define the above parameters is the in vivo testing, implanting the biomaterial in artificially induced bone injury in experimental animals. This is usually carried out after biophysical and biomechanical characterization of the new material, and assessment of bio-tolerability. This is determined as lack of cytotoxic and inflammatory reaction displayed by cells growing on the biomaterial. No further characterization of biomaterial/cells interaction are conducted in vitro, due to the unavailability of adequate cellular model. The proposed cellular model is suitable for the in vitro characterization of osteoinductive and osteogenic properties of biomaterials. We genetically modified a tumor derived cell line, which has been shown to maintain the osteoblastic phenotype and to posses unlimited growing potential in vitro. By stable transfection of an expression vector, containing as reporter gene the enhanced green fluorescent protein (EGFP), we selected an immortal, genetically uniform clonal cell line, the Saos-EGFP. The engineered cell line Saos-EGFP maintains the histological features of osteoblast cells and constitutively expresses the EGFP. The presence of the heterologous protein does not perturb the physiological expression of molecular markers specific of the bone tissue. Comparative analysis carried out on parental and engineered cells demonstrated that both cell populations express molecular markers specific for bone tissue, such as osteocalcin and osteonectin, which belong to the extra cellular matrix proteins, necessary for bone tissue formation, produced by the osteoblasts. Furthermore, the distribution and cellular localization of the marker proteins were undistinguishable between parental and Saos-EGFP cells, indicating that functional properties were not altered. Another criteria of paramount importance in bone tissue histology is the cytoskeletal organization of cells. Studies have shown that the actin cytoskeletal element plays an important role in cell attachment and stabilization. Actin bundles coupled with adhesion plaques can transmit forces to the substrate and help to maintain cell shape and facilitate cell adhesion. Cytoplasmic actin fibers organization was not affected by the presence of the exogenous protein, since a prefect match in the cytoskeletal architecture between the two cell populations was observed. It was shown that the measure of EGFP fluorescence is proportional to the EGFP molecule number within each sample. This means that, having a genetically uniform clonal cell line, the fluorescence intensity is proportional to the number of cell within each sample. The high expression level of the trans-gene in the genetically modified cell line Saos-EGFP ensures a direct proportion between fluorescence intensity and cell number down to 5x103 cells per sample. This relationship is very useful to assess easily and quickly the number of cells within any sample, just determining the fluorescence intensity by a spectrophotometric reading. Thank to the immortal phenotype, the proposed cell line can be mantanined indefinitelly using standard cell culture condition