Identification of appropriate housekeeping genes for quantitative RT-PCR analysis in MDA-MB-231 and NCI-H460 human cancer cell lines under hypoxia and serum deprivation

Gene expression studies aimed at analyzing cancer cells under hypoxia and serum deprivation conditions show major potential for understanding molecular mechanisms associated with tumor progression as well as resistance to antitumor agents. To the best of our knowledge, a study for the identification of appropriate housekeeping genes in breast and lung cancer cells under hypoxia and serum deprivation conditions is currently missing. Given the relevance of a reliable and accurate normalization, we herein aimed to identify the appropriate housekeeping genes for breast and lung cancer cell lines cultured under hypoxia and/or serum deprivation. The stability of five commonly used housekeeping genes (ACTB, β 2M, GUSB, 18S rRNA, and PPIA) was assessed after reverse-transcription quantitative real-time PCR in MDA-MB-231 and NCI-H460 cancer cell lines using GeNorm, NormFinder and BestKeeper software. GeNorm and NormFinder ranking revealed ACTB, GUSB and PPIA as the most stable genes for both tumor cell lines. Our results support the use of ACTB/PPIA for MDA-MB-231 and GUSB/PPIA for NCI-H460 cells as the most stable combination for normalization of gene expression under hypoxic and serum deprivation conditions. Our results highlight the importance of the selection of the housekeeping genes in cancer cells subjected to different physiological stresses, such as hypoxia and serum deprivation.This work was supported by the Coordenação de Aperfeiçoamento de Pessoal de Nıvel Superior (CAPES) [grant agreement No. 88881.132780/2016-01]; Fundac ão de Amparoa Ciência e Tecnologia de Pernambuco (FACEPE) [grant agreement No. 0883-2.08/13];and the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie [grant agreement No. 748880]. This work was also funded by the project NORTE-01-0145-FEDER-000029, supported by Norte Portugal Regional Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF); and by FEDER—Fundo Europeu de Desenvolvimento Regional funds through COMPETE 2020—Operational Programme for Competitiveness and Internationalization (POCI), Portugal 2020,and by Portuguese funds through FCT (Fundação para a Ciência e a Tecnologia)/Ministério da Ciência, Tecnologia e Inovação in the framework of the project “Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274), and the project POCI-01-0145-FEDER-016585 (PTDC/BBB EBI/0567/2014)

Use of mixed methods designs in substance research: a methodological necessity in Nigeria

The utility of mixed methods (qualitative and quantitative) is becoming increasingly accepted in health sciences, but substance studies are yet to substantially benefit from such utilities. While there is a growing number of mixed methods alcohol articles concerning developed countries, developing nations are yet to embrace this method. In the Nigerian context, the importance of mixed methods research is yet to be acknowledged. This article therefore, draws on alcohol studies to argue that mixed methods designs will better equip scholars to understand, explore, describe and explain why alcohol consumption and its related problems are increasing in Nigeria. It argues that as motives for consuming alcohol in contemporary Nigeria are multiple, complex and evolving, mixed method approaches that provide multiple pathways for proffering solutions to problems should be embraced

Housing supply and brownfield regeneration in a post-Barker review world: A comparison of policy and practice in England and Scotland

The findings of the Barker review, which examined the reasons for the undersupply of UK housing, have important implications for the devolved constituents of the UK, including Scotland. This paper traces the emergence of the brownfield regeneration policy agenda across the UK and examines how the Barker review connects with this brownfield policy focus. The paper compares housing and brownfield policies and practices in England and Scotland, places them in an international context and elicits wider lessons for devolved governance in relation to housing policy, in terms of `centrist—local' tensions. Estimates based on published data suggest that Barker's emphasis on increased housing supply cannot easily be reconciled with the current emphasis on brownfield development and is likely to require a return to greenfield development in both countries

Diffusion and adsorption of hydrocarbons from automotive engine exhaust in zeolitic adsorbents

Diffusion characteristics of hydrocarbons typical of automotive engine exhaust were determined by the Zero Length Column (ZLC) chromatographic method in silicalite and ferrierite pellets. The data clearly show that intracrystalline diffusion is too fast to exert any significant influence on the sorption kinetics in adsorbent pellets, and the dominance of meso/macropore diffusional resistance was confirmed by replicate experiments with different particle size fractions. Surface diffusion makes a significant contribution to the meso/macropore diffusivity for toluene in silicalite pellets leading to large differences in effective diffusivities relative to other sorbates, in particular ethylene. Ethylene, iso-butane and iso-butene adsorption and diffusion in ferrierite pellets showed a peculiar behavior due to immobilization effects involving both zeolite crystals and possibly mesoporous walls of the composite particles. At low concentration levels sorbate species diffused as single species in their respective binary mixture

Fine-grained multi-instance classification in microscopy through deep attention

Fine-grained object recognition and classification in biomedical images poses a number of challenges. Images typically contain multiple instances (e.g. glands) and the recognition of salient structures is confounded by visually complex backgrounds. Due to the cost of data acquisition or the limited availability of specimens, data sets tend to be small. We propose a simple yet effective attention based deep architecture to address these issues, specially to improve background suppression and recognition of important instances per image. Attention maps per instance are learnt in an end-to-end fashion. Microscopic images of fungi (new data) and a publicly available Breast Cancer Histology benchmark dataset are used to demonstrate the performance of the proposed approach. Experimental results suggest that the proposed approach advances the state-of-the-art

Optimizing the separation of gaseous enantiomers by simulated moving bed and pressure swing adsorption

The resolution of racemic gas mixtures by simulated moving bed (SMB) and pressure swing adsorption (PSA) is investigated by dynamic simulation and optimization. Enantiomer separation of inhalation anesthetics is important because there is evidence that the purified enantiomers may have different pharmacological properties than the racemate. The model parameters reported in an experimental investigation performed elsewhere are used to study the feasibility of this separation using SMB and PSA configurations. Both processes were modeled in gPROMS® as systems of differential algebraic equations. Operating conditions are optimized such that the feed throughput and product recovery for each process were maximized subject to equal constraints on the pressures and superficial gas velocities. SMB was found to be capable of resolving racemic feed mixtures with purity and recovery exceeding 99%. On the other hand, PSA was also able to provide a single purified enantiomer with low recovery of about 30% which may limit its application to enantiomer separation. Nevertheless, PSA consumes less desorbent, and achieves higher throughput at the sacrifice of lower recovery. © Springer Science+Business Media, LLC 2010 [accessed February 8th 2011

Adsorption and diffusion of propane and propylene in Ag+-impregnated MCM-41

Equilibrium data and diffusion characteristics of propane and propylene were determined on mesoporous adsorbents modified with an organic molecule (APTES) and/or impregnated with AgNO3, in order to obtain a separation by adsorption via π-complexation. Adsorption capacities were determined by a gravimetric technique, while diffusion characteristics were evaluated by the ZLC technique. The equilibrium isotherms data showed that the modification with an organic molecule will weaken the π-interaction between Ag+ and double C=C bond. On the other hand slightly higher adsorption capacities for propylene (about 1.5 mol/kg) were obtained for the sample prepared by a direct impregnation with larger amounts of AgNO3 (M4 sample). Diffusion runs confirmed that the propane desorption rate on M4 sample was much higher compared to propylene. This evidence leads to a potential application of that adsorbent material for a kinetic separation