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.</p

New triazine bridged triads based on BODIPY-porphyrin systems: extended absorption, efficient energy transfer and upconverted emission

Two novel triads connecting a BODIPY to ethylenediamine substituted porphyrins via triazine linker have been synthesized and characterized. One of the triads is a linear D-A structure with one BODIPY (D) and one porphyrin (A) bridged by the triazine linker and the other one is a branched A-D4 structure with the porphyrin core linked to four BODIPY units. The triads show extended absorption in the visible region with contributions from both porphyrin (Soret band centred at 410–430 nm) and BODIPY units (strong absorption at ≈ 502 nm) in good agreement with the expected molar ratio. Both triads exhibit linear and nonlinear optical properties featuring an efficient energy transfer from the BODIPY donor to the porphyrin acceptor. The nonlinear upconverted emission properties of the triads were studied by two-photon excitation in the Near-infrared (NIR, 710–930 nm). The maximum two-photon absorption cross-section values for the triads (40–70 GM) are larger than those typically reported in this wavelength range for porphyrins and BODIPY. Both the green emission of BODIPY (≈514 nm) and the red emission of porphyrins (650–750 nm) were observed under NIR excitation at 930 nm. The distinct features of triads, namely i) an extended absorption; ii) an efficient energy transfer and iii) the nonlinear upconverted emission featuring a large separation between the excitation and emission wavelengths could be beneficial for application in sensing and imaging procedures.publishe

Water and otolith chemistry: implications for discerning estuarine nursery habitat use of a juvenile flatfish

Variations in otolith elemental composition are widely used to reconstruct fish movements. However, reconstructing habitat use and environmental histories of fishes within estuaries is still a major challenge due to the dynamic nature of these coastal environments. In this study, we performed a laboratory experiment to investigate the effects of variations in salinity (three levels; 5, 18, 30) and temperature (two levels; 16, 21 C) on the otolith elemental composition (Mg:Ca, Mn:Ca, Sr:Ca, Ba:Ca) of juvenile Senegalese sole Solea senegalensis. Temperature and salinity treatments mirrored the natural conditions of the estuarine habitats occupied by juvenile Senegalese sole, thereby providing information on the applicability of otolith microchemistry to reconstruct habitat use patterns within estuarine nurseries, where individual fish move across complex salinity and temperature gradients. While Sr:Ca and Ba:Ca in otoliths were both positively related to salinity, no temperature effect was observed. Partition coefficients, proxies for element incorporation rates increased with increasing salinity for Sr (DSr) and Ba (DBa). In contrast, salinity and temperature had little influence on otolith Mn:Ca and Mg:Ca, supporting physiological control on the incorporation of these elements. Our results are a stepping stone for the interpretation of otolith chemical profiles for fish collected in their natural habitats and contribute to better understanding the processes involved in otolith element incorporation.info:eu-repo/semantics/publishedVersio

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.This work was partially supported by the Portuguese Fundacao para a Ciencia e Tecnologia (grant PTDC/SAU-TOX/114549/2009). Olga Martinho and Sara Granja were recipients of PhD fellowships (SFRH/BD/36463/2007 and SFRH/BD/51062/2010, respectively), and Filipe Pinto and Vera Miranda-Goncalves were recipients of research fellowships (UMINHO/BI/016/2011 and SFRH/BI/33503/2008, respectively), both from FCT, Portugal. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. No additional external funding received for this study

Significance of glycolytic metabolism-related protein expression in colorectal cancer, lymph node and hepatic metastasis

Background: Colorectal cancer (CRC) is one of the most common malignancies and a leading cause of cancer death worldwide. Most cancer cells display high rates of glycolysis with production of lactic acid, which is then exported to the microenvironment by monocarboxylate transporters (MCTs). The main aim of this study was to evaluate the significance of MCT expression in a comprehensive series of primary CRC cases, lymph node and hepatic metastasis. Methods: Expressions of MCT1, MCT4, CD147 and GLUT1 were studied in human samples of CRC, lymph node and hepatic metastasis, by immunohistochemistry. Results: All proteins were overexpressed in primary CRC, lymph node and hepatic metastasis, when compared with non-neoplastic tissue, with exception of MCT1 in lymph node and hepatic metastasis. MCT1 and MCT4 expressions were associated with CD147 and GLUT1 in primary CRC. These markers were associated with clinical pathological features, reflecting the putative role of these metabolism-related proteins in the CRC setting. Conclusion: These findings provide additional evidence for the pivotal role of MCTs in CRC maintenance and progression, and support the use of MCTs as biomarkers and potential therapeutic targets in primary and metastatic CRC.This work was supported by the Fundação para a Ciência e a Tecnologia (FCT) grant ref. PTDC/SAU-FCF/104347/2008, under the scope of ‘Programa Operacional Temático Factores de Competitividade’ (COMPETE) of ‘Quadro Comunitário de Apoio III’ and co-financed by the Fundo Europeu De Desenvolvimento Regional (FEDER). Ricardo Amorim was recipient of the fellowship SFRH/BD/98002/2013, from Fundação para a Ciência e a Tecnologia (FCT Portugal).info:eu-repo/semantics/publishedVersio

A voz dos bandos: colectivos de justiça e ritos da palavra portuguesa em Timor Leste colonial

Este artigo examina as relações entre o discurso da justiça e a prática do ritual nos bandos do governo colonial português em Timor Leste, entre a segunda metade do século XIX e as primeiras décadas do século XX. Os bandos consistiam em ordens e instruções de comando emanadas pelo governador português em Díli, e comunicadas de forma cerimonial por oficiais às populações dos diversos reinos timorenses dispersos pelo país. Bandos eram um instrumento por excelência de governação colonial dos assuntos indígenas, servindo para arbitrar conflitos, punir transgressões e, em geral, instituir realidades no mundo timorense. Contudo, esta instituição assumiu igualmente uma singular expressão nos usos timorenses, servindo bandos para comunicar também as ordens de autoridades tradicionais, os liurais. O artigo acompanha as variações coloniais e indígenas que os bandos adquiriram em Timor Leste, conceptualizando-os enquanto colectivos de justiça. Ao considerar assim os bandos como colectivos – formações heterogéneas em que elementos linguísticos e não linguísticos se combinam na produção de efeitos de poder sobre as populações – o artigo propõe uma via conceptual alternativa às perspectivas linguísticas e literárias de análise do discurso colonial

Does an invasive bivalve outperform its native congener in a heat wave scenario? A laboratory study case with Ruditapes decussatus and R. philippinarum

Global warming and the subsequent increase in the frequency of temperature anomalies are expected to affect marine and estuarine species’ population dynamics, latitudinal distribution, and fitness, allowing non-native opportunistic species to invade and thrive in new geographical areas. Bivalves represent a significant percentage of the benthic biomass in marine ecosystems worldwide, often with commercial interest, while mediating fundamental ecological processes. To understand how these temperature anomalies contribute to the success (or not) of biological invasions, two closely related species, the native Ruditapes decussatus and the introduced R. philippinarum, were exposed to a simulated heat wave. Organisms of both species were exposed to mean summer temperature (~18 C) for 6 days, followed by 6 days of simulated heat wave conditions (~22 C). Both species were analysed for key ecological processes such as bioturbation and nutrient generation—which are significant proxies for benthic function and habitat quality—and subcellular biomarkers—oxidative stress and damage, and energetic metabolism. Results showed subcellular responses to heat waves. However, such responses were not expressed at the addressed ecological levels. The subcellular responses to the heat wave in the invasive R. philippinarum pinpoint less damage and higher cellular energy allocation to cope with thermal stress, which may further improve its fitness and thus invasiveness behaviour.info:eu-repo/semantics/publishedVersio

Climate influence on juvenile European sea bass (Dicentrarchus labrax, L.) populations in an estuarine nursery: A decadal overview

Estuarine systems support the life cycle stages of commercially important marine fish and are influenced by large and local-scale climatic patterns. In this study, performed in the Mondego estuary, Portugal, we used an 11-year database (2003-2013) for analyzing the variability in the population of a marine juvenile migrant fish, the European sea bass Dicentrarchus labrax, regarding changes in abundance, population structure, growth rates, secondary production and annual day of peak abundance. Higher densities and production occurred in 2003, but no differences in 0-group growth could be observed. In order to detect change points in both biological and climatic data, the cumulative sum (CUSUM) of the deviations from the mean for the 2003-2013 period were determined for each parameter. The relationship between large and local-scale drivers and 0-group biological attributes were evaluated using a Spearman rank correlation analysis of CUSUM of biological and environmental data, considering the correspondent yearly values and with a time-lag of 1 year. The North Atlantic Oscillation (NAO) index, sea surface temperature (SST) and their respective winter values were tested as large-scale factors, while river runoff, salinity and estuarine water temperature were considered as local climate patterns. The significant factors explaining D. labrax 0-group abundance and production were salinity and the NAO, the latter being also a significant predictor considering the 1-year lag. The NAO with 1-year lag was also positively correlated with the day of peak abundance. The early stages of European sea bass were demonstrated to have a climate-dependent life cycle, controlled by variations in both large-scale climatic patterns and local features. In southern European marine populations, the effects of the NAO seem less direct, and dependent on the magnitude of its expressions and on the time scale considered.info:eu-repo/semantics/publishedVersio

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.This work was supported by the Portuguese Fundacao para a Ciencia e Tecnologia (grant PTDC/SAU-OBD/100079/2008 to P.J., grants PTDC/SAU-IMU/110303/2009 and PTDC/SAU-MII/100780/2008 to L. F. M. and PEst-OE/BIA/UI4046/2011 to the BioFig research unit; fellowships SFRH/BD/36463/2007 to O.M. and UMINHO/BI/016/2011 to F. P.). L. F. M. receives support from Fundacao Luso-Americana para o Desenvolvimento and C.J. acknowledges NHS funding to the Biomedical Research Centre