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This article aims to describe contributions that the Ludological Analysis can offer to the development of serious games. We describe a case study evaluating the characteristics of different digital Serious Games for the understanding of the main elements that make players feel challenged and motivated. This study was conducted based on the application of concepts of Ludology, seeking to validate the factors associated with the liminality, a psychological state in which the player feels so engaged in the gaming experience that focuses, almost entirely, in the game. After identifying these factors in the games analyzed in the case study, these features are mapped to the needs and possibilities in the field of education, specifically in the analysis and planning of digital Serious Games, considering that it's observation may represent improvement in the conception and use of these materials.

The existence of ‘gatekeepers’ presupposes the validity of rules, norms, and laws. Are there any laws in the arts? Many leading art institutions (e.g. academies, conservatories, universities, culture ministries) resist all change, due to their inertia, rather than any explicit defense of principles. The boundaries of art history have become problematic since the seventies, at least; philosophical aesthetics has renounced any claim to be normative; and art criticism is disappearing, to the benefit of curatorship. Perhaps this is the symptom of a deep historical and anthropological change. Is the alleged and practiced ‘creativity’ in the arts (or even in the economy and in life itself) not the opposite of any established norm? Unlike the first sixty years of the twentieth-century – when the ‘cultural industries’ provided pre-packaged ‘arts’, domesticating the taste of the ‘masses’ and allowing people to enjoy short moments of rest before resuming their work – nowadays the popular consumption of artistic and cultural content has become part of the production process. The ‘new’, ‘creative’ and ‘knowledge-based’ economy is bringing – as the main productive power – the most intimate, spiritual resources of shared, single lives into play within production. Economic innovation converges on the artistic avant-garde obligation to produce ever-new ‘languages’. We are benefiting from an unprecedented improvement of the flow of information and of means of production. The individual gaze and ear are directed towards a rapidly growing past. But life is continuously under pressure from a compulsory voracity, and hostage to this ‘cognitive bioeconomy’. However, the process of destruction of the old rules may suggest another opportunity: the opportunity of seeing in the arts gestures that can help us to recognize and free what is single and shared in life. Everyone can be a ‘gatekeeper’, by taking care of what cannot be governed and exploited by that power. This opportunity opens up a different perspective with regard to the arts – and life: a contingent, historical perspective, which nonetheless lies beyond all historicism and the avant-garde dialectic between rules and exceptions.

Extracellular vesicles (EVs) have been shown to play an important role in intercellular communication as carriers of DNA, RNA and proteins. While the intercellular transfer of miRNA through EVs has been extensively studied, the stability of extracellular miRNA (ex-miRNA) once engulfed by a recipient cell remains to be determined. Here, we identify the ex-miRNA-directed phenotype to be transient due to the rapid decay of ex-miRNA. We demonstrate that the ex-miR-223-3p transferred from polymorphonuclear leukocytes to cancer cells were functional, as demonstrated by the decreased expression of its target FOXO1 and the occurrence of epithelial-mesenchymal transition reprogramming. We showed that the engulfed ex-miRNA, unlike endogenous miRNA, was unstable, enabling dynamic regulation and a return to a non-invasive phenotype within 8 h. This transient phenotype could be modulated by targeting XRN1/PACMAN exonuclease. Indeed, its silencing was associated with slower decay of ex-miR-223-3p and subsequently prolonged the invasive properties. In conclusion, we showed that the 'steady step' level of engulfed miRNA and its subsequent activity was dependent on the presence of a donor cell in the surroundings to constantly fuel the recipient cell with ex-miRNAs and of XRN1 exonuclease, which is involved in the decay of these imported miRNA., (© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Gene targeting by microRNAs is important in health and disease. We developed a functional assay for identifying microRNA targets and applied it to the K(+) channel K(ir)2.1 [KCNJ2 (potassium inwardly-rectifying channel, subfamily J, member 2)] which is dysregulated in cardiac and vascular disorders. The 3'UTR (untranslated region) was inserted downstream of the mCherry red fluorescent protein coding sequence in a mammalian expression plasmid. MicroRNA sequences were inserted into the pSM30 expression vector which provides enhanced green fluorescent protein as an indicator of microRNA expression. HEK (human embryonic kidney)-293 cells were co-transfected with the mCherry-3'UTR plasmid and a pSM30-based plasmid with a microRNA insert. The principle of the assay is that functional targeting of the 3'UTR by the microRNA results in a decrease in the red/green fluorescence intensity ratio as determined by automated image analysis. The method was validated with miR-1, a known down-regulator of K(ir)2.1 expression, and was used to investigate the targeting of the K(ir)2.1 3'UTR by miR-212. The red/green ratio was lower in miR-212-expressing cells compared with the non-targeting controls, an effect that was attenuated by mutating the predicted target site. miR-212 also reduced inward rectifier current and K(ir)2.1 protein in HeLa cells. This novel assay has several advantages over traditional luciferase-based assays including larger sample size, amenability to time course studies and adaptability to high-throughput screening.

HL-1 is a line of immortalized cells of cardiomyocyte origin that are a useful complement to native cardiomyocytes in studies of cardiac gene regulation. Several types of ion channel have been identified in these cells, but not the physiologically important inward rectifier K(+) channels. Our aim was to identify and characterize inward rectifier K(+) channels in HL-1 cells. External Ba(2+) (100 µM) inhibited 44 ± 0.05% (mean ± s.e.m., n = 11) of inward current in whole-cell patch-clamp recordings. The reversal potential of the Ba(2+)-sensitive current shifted with external [K(+)] as expected for K(+)-selective channels. The slope conductance of the inward Ba(2+)-sensitive current increased with external [K(+)]. The apparent Kd for Ba(2+) was voltage dependent, ranging from 15 µM at -150  mV to 148 µM at -75  mV in 120  mM external K(+). This current was insensitive to 10 µM glybenclamide. A component of whole-cell current was sensitive to 150 µM 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), although it did not correspond to the Ba(2+)-sensitive component. The effect of external 1 mM Cs(+) was similar to that of Ba(2+). Polymerase chain reaction using HL-1 cDNA as template and primers specific for the cardiac inward rectifier K(ir)2.1 produced a fragment of the expected size that was confirmed to be K(ir)2.1 by DNA sequencing. In conclusion, HL-1 cells express a current that is characteristic of cardiac inward rectifier K(+) channels, and express K(ir)2.1 mRNA. This cell line may have use as a system for studying inward rectifier gene regulation in a cardiomyocyte phenotype., (© 2010 Wiley-Liss, Inc.)

Surface-spread spermatocytes of the fallow deer were examined by light microscopy after silver staining. The development and behaviour of synaptonemal complexes and the partial synapsis of the X and Y chromosomes were investigated during leptotene to diplotene chromosome pairing.

The treatment of termite male spermatogonia with actinomycin D induces highly elongated and finely banded late prophase and prometaphase chromosomes as evidenced by the silver staining method. Actinomycin D suppresses the silver staining of nucleolar organizing regions in prometaphase and reduces it in metaphase chromosomes.