Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells

Bone is constantly being molded and shaped by the action of osteoclasts and osteoblasts. A proper equilibrium between both cell types metabolic activities is required to ensure an adequate skeletal tissue structure, and it involves resorption of old bone and formation of new bone tissue. It is reported that treatment with antiepileptic drugs (AEDs) can elicit alterations in skeletal structure, in particular in bone mineral density. Nevertheless, the knowledge regarding the effects of AEDs on bone cells are still scarce. In this context, the aim of this study was to investigate the effects of five different AEDs on human osteoclastic, osteoblastic and co-cultured cells. Osteoclastic cell cultures were established from precursor cells isolated from human peripheral blood and were characterized for tartrate-resistant acid phosphatase (TRAP) activity, number of TRAP+ multinucleated cells, presence of cells with actin rings and expressing vitronectin and calcitonin receptors and apoptosis rate. Also, the involvement of several signaling pathways on the cellular response was addressed. Osteoblastic cell cultures were obtained from femur heads of patients (25-45 years old) undergoing orthopaedic surgery procedures and were then studied for cellular proliferation/viability, ALP activity, histochemical staining of ALP and apoptosis rate. Also the expression of osteoblast-related genes and the involvement of some osteoblastogenesis-related signalling pathways on cellular response were addressed. For co-cultured cells, osteoblastic cells were firstly seeded and cultured. After that, PBMC were added to the osteoblastic cells and co-cultures were evaluated using the same osteoclast and osteoblast parameters mentioned above for the corresponding isolated cell. Cell-cultures were maintained in the absence (control) or in the presence of different AEDs (carbamazepine, gabapentin, lamotrigine, topiramate and valproic acid). All the tested drugs were able to affect osteoclastic and osteoblastic cells development, although with different profiles on their osteoclastogenic and osteoblastogenic modulation properties. Globally, the tendency was to inhibit the process. Furthermore, the signaling pathways involved in the process also seemed to be differently affected by the AEDs, suggesting that the different drugs may affect osteoclastogenesis and/or osteoblastogenesis through different mechanisms. In conclusion, the present study showed that the different AEDs had the ability to directly and indirectly modulate bone cells differentiation, shedding new light towards a better understanding of how these drugs can affect bone tissue.

Expanding our therapeutic options: β-blockers for colon cancer?

Supported by U. Porto/Santander Totta (IJUP) (PP-IJUP2011-320)

Enoxacin inhibits growth of prostate cancer cells and effectively restores microRNA processing

Prostate cancer (PCa) is one of the most incident malignancies worldwide. Although efficient therapy is available for early-stage PCa, treatment of advanced disease is mainly ineffective and remains a clinical challenge. microRNA (miRNA) dysregulation is associated with PCa development and progression. In fact, several studies have reported a widespread downregulation of miRNAs in PCa, which highlights the importance of studying compounds capable of restoring the global miRNA expression. The main aim of this study was to define the usefulness of enoxacin as an anti-tumoral agent in PCa, due to its ability to induce miRNA biogenesis in a TRBP-mediated manner. Using a panel of five PCa cell lines, we observed that all of them were wild type for the TARBP2 gene and expressed TRBP protein. Furthermore, primary prostate carcinomas displayed normal levels of TRBP protein. Remarkably, enoxacin was able to decrease cell viability, induce apoptosis, cause cell cycle arrest, and inhibit the invasiveness of cell lines. Enoxacin was also effective in restoring the global expression of miRNAs. This study is the first to show that PCa cells are highly responsive to the anti-tumoral effects of enoxacin. Therefore, enoxacin constitutes a promising therapeutic agent for PCa.

Anti-tumoral effect of the non-nucleoside DNMT inhibitor RG108 in human prostate cancer cells

Background: Current therapeutic strategies for advanced prostate cancer (PCa) are largely ineffective. Because aberrant DNA methylation associated with inappropriate gene-silencing is a common feature of PCa, DNA methylation inhibitors might constitute an alternative therapy. In this study we aimed to evaluate the anti-cancer properties of RG108, a novel non-nucleoside inhibitor of DNA methyltransferases (DNMT), in PCa cell lines. Methods: The anti-tumoral impact of RG108 in LNCaP, 22Rv1, DU145 and PC-3 cell lines was assessed through standard cell viability, apoptosis and cell cycle assays. Likewise, DNMT activity, DNMT1 expression and global levels of DNA methylation were evaluated in the same cell lines. The effectiveness of DNA demethylation was further assessed through the determination of promoter methylation and transcript levels of GSTP1, APC and RAR-β2, by quantitative methylation-specific PCR and RT-PCR, respectively. Results: RG108 led to a significant dose and time dependent growth inhibition and apoptosis induction in LNCaP, 22Rv1 and DU145. LNCaP and 22Rv1 also displayed decreased DNMT activity, DNMT1 expression and global DNA methylation. Interestingly, chronic treatment with RG108 significantly decreased GSTP1, APC and RAR-β2 promoter hypermethylation levels, although mRNA re-expression was only attained GSTP1 and APC. Conclusions: RG108 is an effective tumor growth suppressor in most PCa cell lines tested. This effect is likely mediated by reversion of aberrant DNA methylation affecting cancer related-genes epigenetically silenced in PCa. However, additional mechanism might underlie the anti-tumor effects of RG108. In vivo studies are now mandatory to confirm these promising results and evaluate the potential of this compound for PCa therapy.

Phenotypic impact of deregulated expression of class I histone deacetylases in urothelial cell carcinoma of the bladder

Deregulated expression of histone deacetylases (HDACs) has been implicated in tumorigenesis. Herein, we investigated class I HDACs expression in bladder urothelial cell carcinoma (BUCC), its prognostic value and biological significance. Significantly increased transcript levels of all HDACs were found in BUCC compared to 20 normal mucosas, and these were higher in lower grade and stage tumors. Increased HDAC3 levels were associated with improved patient survival. SiRNA experiments showed decrease cell viability and motility, and increased apoptosis. We concluded that class I HDACs play an important role in bladder carcinogenesis through deregulation of proliferation, migration and apoptosis, constituting putative therapeutic targets

Equipamento de produção de água gelada para melhorar a eficácia do combate a incêndios florestais

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Mecânica

Aproveitamento de águas pluviais para rega

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Civil na Área de Especialização de Hidráulica

Projecto de fundações e estrutura de um edifício de habitação utilizando os eurocódigos estruturais

Trabalho de Projecto para obtenção do grau de Mestre em Engenharia Civil Perfil Estruturas

Reabilitação de edifícios com novas tendências NZEB: caso de estudo: edifício de serviços em Setúbal

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Civil

Replica management in real-time Ada 95 applications

In this paper, we present some of the fault tolerance management mechanisms being implemented in the Multi-μ architecture, namely its support for replica non-determinism. In this architecture, fault tolerance is achieved by node active replication, with software based replica management and fault tolerance transparent algorithms. A software layer implemented between the application and the real-time kernel, the Fault Tolerance Manager (FTManager), is the responsible for the transparent incorporation of the fault tolerance mechanisms The active replication model can be implemented either imposing replica determinism or keeping replica consistency at critical points, by means of interactive agreement mechanisms. One of the Multi-μ architecture goals is to identify such critical points, relieving the underlying system from performing the interactive agreement in every Ada dispatching point.

Viabilidade do reforço sísmico de um edifício de pequeno porte em alvenaria de pedra ordinária

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Civil na Área de Especialização de Estruturas

NZEB: um desafio para a engenharia civil

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Civil

Dynamic global scheduling of parallel real-time tasks

High-level parallel languages offer a simple way for application programmers to specify parallelism in a form that easily scales with problem size, leaving the scheduling of the tasks onto processors to be performed at runtime. Therefore, if the underlying system cannot efficiently execute those applications on the available cores, the benefits will be lost. In this paper, we consider how to schedule highly heterogenous parallel applications that require real-time performance guarantees on multicore processors. The paper proposes a novel scheduling approach that combines the global Earliest Deadline First (EDF) scheduler with a priority-aware work-stealing load balancing scheme, which enables parallel realtime tasks to be executed on more than one processor at a given time instant. Experimental results demonstrate the better scalability and lower scheduling overhead of the proposed approach comparatively to an existing real-time deadline-oriented scheduling class for the Linux kernel.

Towards network-on-chip agreement protocols

Demands for functionality enhancements, cost reductions and power savings clearly suggest the introduction of multiand many-core platforms in real-time embedded systems. However, when compared to uni-core platforms, the manycores experience additional problems, namely the lack of scalable coherence mechanisms and the necessity to perform migrations. These problems have to be addressed before such systems can be considered for integration into the realtime embedded domain. We have devised several agreement protocols which solve some of the aforementioned issues. The protocols allow the applications to plan and organise their future executions both temporally and spatially (i.e. when and where the next job will be executed). Decisions can be driven by several factors, e.g. load balancing, energy savings and thermal issues. All presented protocols are analytically described, with the particular emphasis on their respective real-time behaviours and worst-case performance. The underlying assumptions are based on the multi-kernel model and the message-passing paradigm, which constitutes the communication between the interacting instances.

Incorporação de resíduo da indústria petrolífera em argamassas de cal hidráulica para a reabilitação de edifícios

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Civil na Área de Especialização de Edificações