Investigating the role of Sphingosine Kinase 1 pathway in cancer cell-monocyte interactions

Strong evidence suggests that the tumour microenvironment is inflammatory and that activation of the innate immune system plays a role in cancer progression, therefore targeting the multiple interactions of tumour cells with other cell types within the tumour microenvironment may lead to development of new cancer therapies. Sphingosine kinase (SPHK1) is a tumour-associated enzyme whose over-expression has been linked to patient mortality in many types of cancer. Here I investigate whether activation of the SPHK1 pathway, with a known involvement in inflammatory responses, is a signal transduction component of the tumour-monocyte/macrophage cellular interaction and a key element in inflammation-related cancer progression. Using a co-culture model, this study shows that the presence of monocytes increases cancer cell proliferation, an effect abrogated by knockdown of SPHK1 in cancer cells. Both monocytes and cancer cells showed a transient increase in SPHK1 activity and mRNA expression levels together with an increase in MCP-1 and IL-6 secretion. Silencing of SPHK1 in cancer cells abrogated SPHK1 activation in monocytes and pharmacological inhibition of SPHK1 in monocytes cells decreased monocyte induced-SPHK1 expression in cancer cells. Mechanistically, activation of AKT was observed in cancer cells upon co-culture with monocytes, an effect that was abrogated when cancer cells were pre-treated with siRNA for SPHK1. Moreover, the increase of phospho-AKT, ERK1/2 and NF-KB in monocytes by cancer cells was also reduced by RNAi-mediated knockdown of SPHK1 in cancer cells. My data show that STAT1 can bind to SPHK1 promoter or coding region and may be involved in SPHK1 transcriptional regulation in cancer cells upon monocyte stimulation, however its role still remains unclear as it acts as a transcriptional repressor of SPHK1. Monocytes induced cancer cell chemoprotection via a SPHK1-dependent mechanism, and reduced the inhibitory effect of docetaxel on cancer cell proliferation. Accordingly, increased AKT and ERK1/2 phosphorylation in monocytes were also affected by siRNA targeting of SPHK1 in docetaxel treated cancer cells. Altogether I show for the first time that selective inhibition of SPHK1 in tumour cells can affect their interaction with surrounding cells through the modulation of signal transduction pathways (ERK, PI3K, NF-kB) and cytokine exchange (IL-6, MCP-1 and potentially S1P, GM-CSF, GROα, IL-32 and ICAM-1). SPHK1-mediated increase in proliferation and chemoresistance of cancer cells conferred by monocytes renders this enzyme a promising target for future cancer therapies.Open Acces

BALANCE 4P: Balancing decisions for urban brownfield redevelopment. Technical report of the BALANCE 4P project of the SNOWMAN Network coordinated call IV.

Land take as a result of urbanization is one of the major soil threats in Europe. One of the key measures to prevent further urban sprawl and additional land take, is redevelopment of urban brownfields: underused urban areas with, in many cases, soil and groundwater pollution. The latter issue can be a bottleneck for redevelopment of brownfields instead of green fields. A difficulty for brownfield redevelopments is that in urban projects the responsibilities, tools and knowledge of subsurface engineering and urban planning and design are not integrated; they depend heavily on each other but work in sectors. The urban designer usually deals with opportunities for socio-economic benefits while the subsoil engineer deals with the technical challenges of the site. Balance 4P suggests a holistic approach to brownfield redevelopment that (i) recognizes all phases of the urban redevelopment process which are influenced by the planning conditions set by laws, regulations, policy and institutions; (ii) acknowledges multiple subsurface qualities in the brownfield redevelopment project; (iii) promotes knowledge exchange between the surface and the subsurface sectors, across disciplines within each sector, and over time, about the subsurface qualities of the specific project; (iv) focus on the urban redevelopment project by identifying strategies for redevelopment that can fulfil a good quality of the built environment; (v) assesses the three P’s (People, Planet, Profit/Prosperity) in each urban redevelopment phase; and (vi) puts the Process in focus rather than specific instruments by focusing on identification of WHO should be involved in the knowledge exchange process and HOW it can be mediated. The developed decision support framework is aimed to guide project teams willing to implement a more holistic approach in practice. The framework includes four steps carried out in iterative manner: (1) stakeholder analysis, (2) generation of redevelopment alternatives, (3) sustainability assessment of the alternatives, and (4) synthesis of the assessment results, including uncertainty analysis. The guidance describing the steps in the decision support framework and activities within each step can help to structure the decision process and provide support to project teams. The anticipated advantages of the holistic approach are redevelopment plans that allow for smart, cost-effective and sustainable solutions in the implementation process by making explicit use of subsurface information and knowledge in the planning process, and possibilities for more long-term sustainable planning with regard to the subsurface by increased awareness of the subsurface as a resource and the associated risks and possibilities

Desenvolvimento de modelo lagrangiano de partículas considerando os efeitos do vento e espalhamento de manchas de óleo

A modelagem computacional é uma importante ferramenta para estimar a trajetória e destino final de manchas de óleo em diferentes condições ambientais, visto a complexidade dos processos que atuam nesse poluente. O presente trabalho concentrou os esforços no desenvolvimento de um modelo lagrangiano de trajetória de partículas que simule o movimento de manchas de óleo em ambiente marinho. O modelo utilizado é o Modelo Lagrangiano de Partículas com Deslocamento Aleatório (MLPDA), que é baseado na equação de Langevin. Em princípio, o algoritmo da advecção da mancha de óleo devido ao vento é implementado no MLPDA, visto sua importância ao deslocamento das partículas. É considerado que 3% da velocidade do vento a 10 metros de altura permite uma boa representação da deriva de manchas de óleo em ambiente marinho. Os testes para este algoritmo apresentaram resultados satisfatórios. Posteriormente, é implementado um algoritmo que representa o processo físico de espalhamento do óleo, conhecido também por espalhamento mecânico, que é definido como o movimento horizontal devido às forças gravitacionais, viscosas e inerciais. No presente estudo, esse processo é fundamentando nas equações definidas por Lehr et al. (1984), onde os resultados dos testes mostraram que as partículas espalham conforme exposto por esse mesmo autor e são influenciadas até cerca de 100 h de simulação. Ainda neste estudo, é avaliado o módulo de cálculo de área implementado no MLPDA. É advertido que malhas grosseiras podem resultar em áreas superestimadas, sendo aconselhável o uso de malhas mais refinadas para o cálculo dessas áreas. Por fim, três cenários de simulação de um derrame hipotético de óleo na Baía do Espírito Santo, no interior do Porto de Tubarão, são conduzidos para ilustrar uma aplicação do modelo desenvolvido. As simulações expõem que há grandes diferenças entre os resultados obtidos, principalmente entre o cenário que desconsidera o vento e os outros dois com a consideração desta forçante. O primeiro cenário, as partículas tenderam a permanecer na Baía do Espírito Santo, enquanto para os demais cenários as partículas caminharam para os canais do sistema estuarino da Grande Vitória (Canal da Passagem e Canal de Acesso aos Portos)

An investigation of the emmunomodulatory properties of Sutherlandia Frutescens and Hypoxis Hermerocallidea

Human immunodeficiency virus (HIV) is currently the most significant infectious pathogen and the causative agent of acquired immune deficiency syndrome (AIDS). Unfortunately, due to lack of resources, delivery of antiretroviral therapy (ART) to countries where they are most needed, such as South Africa, Botswana, Lesotho, Malawi and Swaziland, is limited and inefficient. Moreover, the short supply and high cost of antiretroviral drugs have caused researchers to turn to plants as prospective therapies in the search for alternative anti-HIV or immunomodulatory compounds. In an African context, traditional medicines are of great importance, not so much as an alternative to treatment, but in many cases as the only source of treatment. There are various South African plants used medicinally which possess phytochemical constituents that target certain mediators of inflammation and the immune system. In African regions where patients do not have access or financial capability to obtain conventional antiretroviral treatment, traditional herbal medicines are used as primary treatment of HIV/AIDS, regardless of the fact that the safety, toxicity and efficacy of these products are not yet fully understood and that a risk for adverse effects exists. Hypoxis hemerocallidea Fisch&C. A. Mey. (Hypoxidaceae) as well as Sutherlandia frutescens L. R. Br. (Leguminosae) have various effects on the immune system and due to claims about their immune boosting properties, they are two of the most common African herbal compounds being used for HIV management in South Africa. In this study, the immune modulating properties of H. hemerocallidea and S. frutescens were investigated in order to determine whether anectodal claims made about these plants could be supported. Differentiated THP-1 and U937 macrophages were treated with aqueous extracts of H. hemerocallidea and S. frutescens as well as with solutions of compound standards reputedly isolated from these plants such as beta-sitosterol, found in H. hemerocallidea, canavanine, pinitol and gammaaminobutyric acid (GABA) which are present in S. frutescens Cytotoxicity of the test compounds was determined using the 3-(4,5-dimethylthiazol- 2-yl)-2,5-dephenyl tetrazolium bromide (MTT) assay. Antioxidant capacity was assessed using the Trolox equivalence antioxidant capacity (TEAC) and Oxygen radical antioxidant capacity (ORAC) assays. Determination of prostaglandin E2 (PGE2) concentration in treated THP-1 and U937 cell culture supernatants was performed by ELISA. Concentrations of cytokines (IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, TNF-á and IFN-ϒ) in treated THP-1 and U937 cell culture supernatants were determined by flow cytometry. Curcumin, a well-known immunomodulatory compound, was used as a positive control. Results of cytotoxicity assessments showed that H. hemerocallidea (0.1 – 1.9 mg/ml), S. frutescens (0.1 – 1.6 mg/ml), beta-sitosterol (0.2 – 25 ìM), canavanine, pinitol and GABA (1.5 – 200 ìM) were not cytotoxic to THP-1 and U937 macrophages and had cytotoxicity profiles comparable to that of the positive control, curcumin (0.8 - 25 ìM). The TEAC and ORAC assays showed different results in the antioxidant capacities of the test compounds. The purported antioxidant activity of H. hemerocallidea was confirmed by the TEAC assay with antioxidant effects equivalent to 0.2 mg/ml Trolox. Canavanine showed antioxidant activity equivalent to approximately 0.17 mg/ml Trolox and comparable to that of curcumin in the ORAC assay, suggesting its involvement in the inhibition of peroxyl radical-induced oxidation. Flow cytometry results showed that curcumin (20 ìg/ml and 10 ìg/ml) and beta-sitosterol (25 ìg/ml and 12.5 ìg/ml) reduced IL-1â and IL-8 production and significantly (p<0.05) decreased the production of TNF-á. This suggests that betasitosterol could indeed possess anti-inflammatory properties, with effects comparable to the known anti-inflammatory effect of curcumin in terms of cytokine profiles. Beta-sitosterol (25 ìg/ml) and pinitol (50 ìg/ml) significantly (p<0.001) decreased extracellular PGE2 levels in U937 macrophages by 233.4 pg/ml and 281.7 pg/ml, respectively and were the only two compounds showing greater reductions in PGE2 than curcumin. In conclusion, results of this study do not provide enough evidence to support all anecdotal claims about the ‘immune boosting’ properties of S. frutescens and H. hemerocallidea, but the compounds canavanine, beta-sitosterol and pinitol were found to have modulatory effects on certain aspects of the immune system.Dissertation (MSc)--University of Pretoria, 2011.Pharmacologyunrestricte

Semantic Harmonization for Seamless Networked Supply Chain Planning in the Future of Internet

International audienceToday, enterprises are typically in a constant process of acquiring and updating its information technologies, however typically without an overall view of the global inter and intra enterprise's system integration. Foreseeing the future of internet, researchers have been proposing new methodologies and platforms to assist such integration in the network of applications and data. However, implementing new technologies in organizations is a difficult task, since its quality needs for architectures development are more exigent and critical than ever, due to the systems complexity and dimension, semantic needs, and to the interoperability requirements to interact with third party applications and infrastructures. Scientific foundations for EI are envisaged to roadmap such general knowledge covering the general laws of the operation on enterprise interoperability in the future of internet. This paper reports research results from ongoing European Commission supported projects that are members of the Future Internet Enterprise Systems (FInES) European Cluster. The paper draws concepts from the complex systems science and proposes a methodology for seamless networked Supply Chain Planning (SCP), by using a domain reference ontology, data model representation standards, software components evaluation and interoperability checking processes. The methodology VALTE is used to assure that enterprises use tools for SCP compliant to semantics, represented in a common reference ontology, created by the MENTOR methodology. These two horizontal methodologies are vertically supported by interoperability checking processes, which contribute for an interoperable supply chain planning system on the future internet

From Social Dialogue to the Just Transition: A case study analysis of the Yorkshire and the Humber region

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CONTRIBUIÇÃO PARA MELHORIA DA GESTÃO DO SERVIÇO PÚBLICO. Estudo de caso: análise da evolução do desempenho do Inmetro

The search for continuous improvement should be intrinsic to all the processes and present in the daily procedures of the organizations that are constantly looking for the implementation of integrated administration through total quality, based on the Approaches of Excellency. In this sense, the present study considers a critical reflection about the effective use of the Evaluation Report of the Federal Government's Quality Award - PQGF, for the National Institute of Metrology, Standardization and Industrial Quality - Inmetro. It treats the period concerning the 1998 award, when the institution has received the silver ribbon award, to conquest lately in 2000 the prize in the gold ribbon category. The dissertation was based in the case study parameters. In this research work the Reports of Administration presented by Inmetro to PQGF and in the Reports of Evaluation guided by the managers of the program to Inmetro, specifically those relative to the cycles of award of 1998 and of 2000, have been used. The theoretical background, was achieved from a bibliographical revision of subjects treated in text Approaches of Excellency of the National Foundation of Quality Award - FPNQ, year 2002, which are the conceptual base of most of the awards implemented in BrazilUniversidade Federal FluminenseA melhoria contínua deve estar intrínseca em todos os processos e no cotidiano das organizações que buscam constantemente a implantação de gestão integrada pela qualidade total, baseada nos Critérios de Excelência. Neste sentido, o presente estudo faz uma reflexão crítica sobre a efetiva utilização do Relatório de Avaliação do Prêmio Qualidade do Governo Federal - PQGF, pelo Instituto Nacional de Metrologia, Normalização e Qualidade Industrial Inmetro, relativo ao ciclo de premiação de 1998, quando a instituição foi avaliada na faixa de premiação prata, para a conquista do prêmio na faixa ouro, no ciclo de 2000. A dissertação foi realizada segundo os parâmetros de estudo de caso, baseando-se em pesquisa nos Relatórios de Gestão apresentados pelo Inmetro ao PQGF e nos Relatórios de Avaliação encaminhados pelos gestores do programa ao Inmetro, especificamente aqueles relativos aos ciclos de premiação de 1998 e de 2000. Os fundamentos teóricos foram estruturados a partir dos Critérios de Excelência da Fundação Prêmio Nacional da Qualidade FPNQ e de uma revisão bibliográfica de cada item, tendo como base o ciclo de 2002. Estes critérios constituem a base conceitual da maioria dos prêmios implementados no Brasi