Filipe Pinto

T-box transcription factor brachyury is associated with prostate cancer progression and aggressiveness.

Purpose: Successful therapy of patients with prostate cancer is highly dependent on reliable diagnostic and prognostic biomarkers. Brachyury is considered a negative prognostic factor in colon and lung cancer; however, there are no reports on Brachyurys expression in prostate cancer. Experimental Design: In this study, we aimed to assess the impact of Brachyury expression in prostate tumorigenesis using a large series of human prostate samples comprising benign tissue, prostate intraepithelial neoplasia (PIN) lesions, localized tumor, and metastatic tissues. The results obtained were compared with what can be inferred from the Oncomine database. In addition, multiple in vitro models of prostate cancer were used to dissect the biologic role of Brachyury in prostate cancer progression. Results: We found that Brachyury is significantly overexpressed in prostate cancer and metastatic tumors when compared with normal tissues, both at protein and at mRNA levels. Brachyury expression in the cytoplasm correlates with highly aggressive tumors, whereas the presence of Brachyury in the nucleus is correlated with tumor invasion. We found that Brachyury-positive cells present higher viability, proliferation, migration, and invasion rates than Brachyury-negative cells. Microarray analysis further showed that genes co-expressed with Brachyury are clustered in oncogenic-related pathways, namely cell motility, cell-cycle regulation, and cell metabolism. Conclusions: Collectively, the present study suggests that Brachyury plays an important role in prostate cancer aggressiveness and points, for the first time, to Brachyury as a significant predictor of poor prostate cancer prognosis. Our work paves the way for future studies assessing Brachyury as a possible prostate cancer therapeutic target. Clin Cancer Res; 20(18); 4949–61. ©2014 AACR.

Loss of WNK2 expression by promoter gene methylation occurs in adult gliomas and triggers Rac1-mediated tumour cell invasiveness

The gene encoding protein kinase WNK2 was recently identified to be silenced by promoter hypermethylation in gliomas and meningiomas, suggesting a tumour-suppressor role in these brain tumours. Following experimental depletion in cell lines, WNK2 was further found to control GTP-loading of Rac1, a signalling guanosine triphosphatase involved in cell migration and motility. Here we show that WNK2 promoter methylation also occurs in 17.5% (29 out of 166) of adult gliomas, whereas it is infrequent in its paediatric forms (1.6%; 1 out of 66). Re-expression of WNK2 in glioblastoma cells presenting WNK2 gene silencing reduced cell proliferation in vitro, tumour growth in vivo and also cell migration and invasion, an effect correlated with reduced activation of Rac1. In contrast, when endogenous WNK2 was depleted from glioblastoma cells with unmethylated WNK2 promoter, changes in cell morphology, an increase in invasion and activation of Rac1 were observed. Together, these results validate the WNK2 gene as a recurrent target for epigenetic silencing in glia-derived brain tumours and provide first mechanistic evidence for a tumour-suppressing role of WNK2 that is related to Rac1 signalling and tumour cell invasion and proliferation.

RKIP Inhibition in cervical cancer Is associated with higher tumor aggressive behavior and resistance to cisplatin therapy

Cervical cancer is one of the most common cancers in women worldwide, being high-risk group the HPV infected, the leading etiological factor. The raf kinase inhibitory protein (RKIP) has been associated with tumor progression and metastasis in several human neoplasms, however its role on cervical cancer is unclear. In the present study, 259 uterine cervix tissues, including cervicitis, cervical intraepithelial lesions and carcinomas, were analyzed for RKIP expression by immunohistochemistry. We found that RKIP expression was significantly decreased during malignant progression, being highly expressed in non-neoplastic tissues (54% of the samples; 73/135), and expressed at low levels in the cervix invasive carcinomas (∼15% (19/124). Following in vitro downregulation of RKIP, we observed a viability and proliferative advantage of RKIP-inhibited cells over time, which was associated with an altered cell cycle distribution and higher colony number in a colony formation assay. An in vitro wound healing assay showed that RKIP abrogation is associated with increased migratory capability. RKIP downregulation was also associated with an increased vascularization of the tumors in vivo using a CAM assay. Furthermore, RKIP inhibition induced cervical cancer cells apoptotic resistance to cisplatin treatment. In conclusion, we described that RKIP protein is significantly depleted during the malignant progression of cervical tumors. Despite the lack of association with patient clinical outcome, we demonstrate, in vitro and in vivo, that loss of RKIP expression can be one of the factors that are behind the aggressiveness, malignant progression and chemotherapy resistance of cervical cancer.

IoT architecture proposal for disabled people

We are living in a new communication age, which will radically transform the way we live in our society. A world where anything will be connected to Internet is being created, generating an entirely new dynamic network - The Internet of Things (IoT) - enabling new means of communication between people, things and environment. A suitable architecture for Internet of Things (IoT) demands the implementation of several and distinct technologies that range from computing (e.g. Cloud Computing), communications (e.g. 6LowPAN, 3/4G) to semantic (e.g. data mining). Thus, it is necessary to understand very well all these technologies in order to know which of them is most suitable for a given scenario. Therefore, this paper proposes an IoT architecture for disabled people and intends to identify and describe the most relevant IoT technologies and international standards for the stack of the proposed architecture. In particular, the paper discusses the enabling IoT technologies and its feasibility for people with disabilities. At the end, it presents two use cases that are currently being deployed for this population.

Brachyury identifies a class of enteroendocrine cells in normal human intestinal crypts and colorectal cancer

Normal homeostasis of adult intestinal epithelium and repair following tissue damage is maintained by a balance of stem and differentiated cells, many of which are still only poorly characterised. Enteroendocrine cells of the gut are a small population of differentiated, secretory cells that are critical for integrating nutrient sensing with metabolic responses, dispersed amongst other epithelial cells. Recent evidence suggests that sub-sets of secretory enteroendocrine cells can act as reserve stem cells. Given the link between cells with stem-like properties and cancer, it is important that we identify factors that might provide a bridge between the two. Here, we identify a sub-set of chromogranin A-positive enteroendocrine cells that are positive for the developmental and cancer-associated transcription factor Brachyury in normal human small intestinal and colonic crypts. Whilst chromogranin A-positive enteroendocrine cells are also Brachyury-positive in colorectal tumours, expression of Brachyury becomes more diffuse in these samples, suggesting a more widespread function in cancer. The finding of the developmental transcription factor Brachyury in normal adult human intestinal crypts may extend the functional complexity of enteroendocrine cells and serves as a platform for assessment of the molecular processes of intestinal homeostasis that underpins our understanding of human health, cancer and aging.

SPINT2 deregulation in prostate carcinoma

SPINT2 is a tumor suppressor gene that inhibits proteases implicated in cancer progression, like HGFA, hepsin and matriptase. Loss of SPINT2 expression in tumors has been associated with gene promoter hypermethylation; however, little is known about the mechanisms of SPINT2 deregulation in prostate cancer (PCa). We aimed to analyze SPINT2 expression levels and understand the possible regulation by SPINT2 promoter hypermethylation in PCa. In a cohort of 57 cases including non-neoplastic and PCa tissues, SPINT2 expression and promoter methylation was analyzed by immunohistochemistry and methylation-specific PCR, respectively. Methylation status of the SPINT2 promoter was also evaluated by bisulfite sequencing and 5-aza-2’-deoxycytidine treatment. Oncomine and TCGA databases were used to perform in silico PCa analysis of SPINT2 mRNA and methylation levels. A reduction in SPINT2 expression levels from non-neoplastic to PCa tissues was observed; however, none of the cases exhibited SPINT2 promoter methylation. Both bisulfite sequencing and 5-aza demonstrated that SPINT2 promoter is not methylated in PCa cells. Bioinformatics approaches did not show downregulation of SPINT2 at the mRNA level and, in corroboration with our results, SPINT2 promoter region is reported to be unmethylated. Our study suggests an involvement of SPINT2 in PCa tumorigenesis, probably in association with a post-translational regulation of SPINT2.

A Conceptual Framework for Marketing Intelligence

Nowadays, customers demand faster customer service, and expect that organizations know them and provide appropriate services and recommendations for products quickly. Many organizations are reacting to these market needs by driving toward pervasive business intelligence, augmenting traditional business intelligence (BI) with the ability to capture, interpret and act immediately on data to make faster decisions. With pervasive business intelligence, organizations data warehouse changes to a system that can proactively and reactively interact with the business stakeholders, and it provides appropriate decision options to help marketers to respond and take action based on knowledge discovery in current integrated data. This work based on the literature review presents the Pervasive Business Intelligence state of the art and related areas, leaving open for proposing a conceptual framework to guide the development of activities of Marketing Intelligence. The analysis of large volumes of data is impossible without resorting to the appropriate software tools, making it essential to develop frameworks that help to automatically and intelligently, analyzing, interpreting and correlating data, enabling the development and selection of strategies for action (2). In order to assist companies in this exploration of data, concepts and tools for organizing information are critical, highlighting the Pervasive Business Intelligence (PBI) and Marketing Intelligence (MKTI) as pillars to support the decision-making. The economic decline is impelling organizations to examine ways of retaining customers, speed up their services, spending less capital be more efficient regarding their budgets, and observing regulations. Business intelligence (BI) is the ability to access data from multiple sources in an organization and deliver it to appropriate business users for analysis. Manage the performance of the business means know what questions to ask and have the facts at hand at time to answer them, and this is what business intelligence delivers. With pervasive business intelligence, organizations data warehouse changes to a system that can proactively and reactively interact with the business stakeholders, and it provides appropriate decision options to help marketers to respond and take action based on knowledge discovery in current integrated data. Pertinent and accurate information from relevant and reliable sources entails to be successfully processed. This implies that a company needs to be confident it has the right information, at the right time, and dissembled to right people (3). We propose a framework to guide the development of activities of Marketing. A framework of satisfying information needs for decision-making is complex and is compound by different activities to be exploited. This paper is organized as follows: after this introductory part we present related background concepts. Then, the main contribution is presented in terms of a framework proposal. Finally we draw some conclusions.

INTCare: On-line knowledge discovery in the intensive care unit

In our work aim to automate the knowledge discovery process. In this paper we present the INTCare system, an intelligent decision support system for intensive care medicine. INTCare is an agent based system that has (autonomous) agents responsible both for data acquisition and model updating thus reducing the need for human intervention. In the present, INTCare is predicting organ failure and probability of in-hospital death. Reliable prediction results facilitate a change from the current reactive behavior to a pro-active one thus enhancing the quality of service. The functional and structural aspects are presented as are some results obtained using data collected from the bedside monitors.

Pervasive business intelligence as a competitive advantage

Today the strategic significance of information is fundamental to any organization. With the intensification of competition between companies in open markets and often saturated, companies must learn to know themselves and to the market through the collection and analysis of quality information. The strategic information is seen as a key resource for success in the business, which is provided by Business Intelligence systems. A successful business strategy requires an awareness of the surrounding (internal and external) environment of organizations, including customers, competitors, industry structure and competitive forces. Managing the future means not only is able to anticipate what will happen outside the organization, but also be able to represent the events through their own actions timely. To make it possible, Pervasive Business Intelligence arises as a natural evolution of business intelligence applications in organizations, allowing to companies achieve and maintain a sustainable competitive advantage.

Penetration Testing on Virtual Environments

Since the beginning, computer systems have faced the challenge of protecting the information with which they work, and with the technological development, computational security techniques have become more complex to face the potentials attacks. Currently we are facing a war game with the usual two sides, attackers and defenders. The attackers want to have complete control over the systems. In is turn, defenders virtualized systems to maintain the resources safety in case of attack. The attackers have also developed increasingly sophisticated techniques to break such protections, being necessary to anticipate such events, which may be achieved through the application of preventative measures. This may be done by simulating Penetration Testing (PT). PT is an attack on a computer system, using a set of specialized tools that looks for security weaknesses, which eventually may have access to computers features and data, allowing the discovery of such evidence vulnerability. Virtual Environments have a higher exposure to cyber-attacks. The aim of this paper is propose a framework to provide guidelines for Penetration Testing in Virtual Environments.