107,332 research outputs found

    Batch production of a silk-elastin-like protein in E. coli BL21(DE3) : key parameters for optimisation

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    The authors wish to thank Prof. Dorit Schuller and InĂȘs Mendes for the use of and assistance with the HPLC column and all the technical staff at the CBMA for their skilful technical assistance.Background: Silk-elastin-like proteins (SELPs) combining the physicochemical and biological properties of silk and elastin have a high potential for use in the pharmaceutical, regenerative medicine and materials fields. Their development for use is however restrained by their production levels. Here we describe the batch production optimisation for a novel recently described SELP in the pET-E. coli BL21(DE3) expression system. Both a comprehensive empirical approach examining all process variables (media, induction time and period, temperature, pH, aeration and agitation) and a detailed characterisation of the bioprocess were carried out in an attempt to maximise production with this system. Results: This study shows that maximum SELP volumetric production is achieved at 37°C using terrific broth at pH 6–7.5, a shake flask volume to medium volume ratio of 10:1 and an agitation speed of 200 rpm. Maximum induction is attained at the beginning of the stationary phase with 0.5 mM IPTG and an induction period of at least 4 hours. We show that the selection agents ampicillin and carbenicillin are rapidly degraded early in the cultivation and that plasmid stability decreases dramatically on induction. Furthermore, acetate accumulates during the bioprocess to levels which are shown to be inhibitory to the host cells. Using our optimised conditions, 500 mg/L of purified SELP was obtained. Conclusions: We have identified the optimal conditions for the shake flask production of a novel SELP with the final production levels obtained being the highest reported to date. While this study is focused on SELPs, we believe that it could also be of general interest to any study where the pET (ampicillin selective marker)-E. coli BL21 (DE3) expression system is used. In particular, we show that induction time is critical in this system with, in contrast to that which is generally believed, optimal production being obtained by induction at the beginning of the stationary phase. Furthermore, we believe that we are at or near the maximum productivity for the system used, with rapid degradation of the selective agent by plasmid encoded ÎČ-lactamase, plasmid instability on induction and high acetate production levels being the principal limiting factors for further improved production. Keywords: Biopolymers, Silk-elastin like polymers, pET-E. coli BL21(DE3), Batch productionThis work was financed by the European Commission, via the 7th Framework Programme Project EcoPlast (FP7-NMP-2009-SME-3, collaborative project number 246176), by FEDER through POFC – COMPETE and by national funds from Fundação para a CiĂȘncia e Tecnologia (FCT) through PEst project C/BIA/UI4050/2011C/BIA/UI4050/2011. AC is a recipient of FCT grant SFRH/ BD/75882/2011. TC is thankful to FCT for its support through Programa CiĂȘncia 2008.Fundação para a CiĂȘncia e a Tecnologia (FCT

    Localism vs. Individualism for the Scientific Realism Debate

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    Localism is the view that the unit of evaluation in the scientific realism debate is a single scientific discipline, sub-discipline, or claim, whereas individualism is the view that the unit of evaluation is a single scientific theory. Localism is compatible, while individualism is not, with a local pessimistic induction and a local selective induction. Asay (2016) presents several arguments to support localism and undercut globalism, according to which the unit of evaluation is the set of all scientific disciplines. I argue that some of his arguments clash with localism as well as with globalism and support individualism, and that individualism goes hand in hand, while localism does not, with the basic rule of how to evaluate an argument

    Specificity and coherence of body representations

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    Bodily illusions differently affect body representations underlying perception and action. We investigated whether this task dependence reflects two distinct dimensions of embodiment: the sense of agency and the sense of the body as a coherent whole. In experiment 1 the sense of agency was manipulated by comparing active versus passive movements during the induction phase in a video rubber hand illusion (vRHI) setup. After induction, proprioceptive biases were measured both by perceptual judgments of hand position, as well as by measuring end-point accuracy of subjects' active pointing movements to an external object with the affected hand. The results showed, first, that the vRHI is largely perceptual: passive perceptual localisation judgments were altered, but end-point accuracy of active pointing responses with the affected hand to an external object was unaffected. Second, within the perceptual judgments, there was a novel congruence effect, such that perceptual biases were larger following passive induction of vRHI than following active induction. There was a trend for the converse effect for pointing responses, with larger pointing bias following active induction. In experiment 2, we used the traditional RHI to investigate the coherence of body representation by synchronous stimulation of either matching or mismatching fingers on the rubber hand and the participant's own hand. Stimulation of matching fingers induced a local proprioceptive bias for only the stimulated finger, but did not affect the perceived shape of the hand as a whole. In contrast, stimulation of spatially mismatching fingers eliminated the RHI entirely. The present results show that (i) the sense of agency during illusion induction has specific effects, depending on whether we represent our body for perception or to guide action, and (ii) representations of specific body parts can be altered without affecting perception of the spatial configuration of the body as a whole

    Selective Realism vs. Individual Realism for Scientific Creativity

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    Individual realism asserts that our best scientific theories are (approximately) true. In contrast, selective realism asserts that only the stable posits of our best scientific theories are true. Hence, individual realism recommends that we accept more of what our best scientific theories say about the world than selective realism does. The more scientists believe what their theories say about the world, the more they are motivated to exercise their imaginations and think up new theories and experiments. Therefore, individual realism better fosters scientific creativity than selective realism

    Metal-organic frameworks as selective or chiral oxidation catalysts

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    Since the discovery of Metal Organic Frameworks (MOFs) in the early 1990s, the amount of new structures has grown exponentially. A MOF typically consists of inorganic nodes that are connected by organic linkers to form crystalline, highly porous structures. MOFs have attracted a lot of attention lately, as the versatile design of such materials holds promises of interesting applications in various fields. In this review, we will focus on the use of MOFs as heterogeneous oxidation catalysts. MOFs are very promising candidates to replace homogeneous catalysts by sustainable and stable heterogeneous catalysts. The catalytic active function can be either the active metal sites of the MOF itself or can be introduced as an extra functionality in the linker, a dopant or a "ship-in-a-bottle" complex. As the pore size, pore shape, and functionality of MOFs can be designed in numerous ways, shape selectivity, and even chiral selectivity can be created. In this article, we will present an overview on the state of the art of the use of MOFs as a heterogeneous catalyst in liquid phase oxidation reactions

    Positive emotion broadens attention focus through decreased position-specific spatial encoding in early visual cortex: evidence from ERPs

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    Recent evidence has suggested that not only stimulus-specific attributes or top-down expectations can modulate attention selection processes, but also the actual mood state of the participant. In this study, we tested the prediction that the induction of positive mood can dynamically influence attention allocation and, in turn, modulate early stimulus sensory processing in primary visual cortex (V1). High-density visual event-related potentials (ERPs) were recorded while participants performed a demanding task at fixation and were presented with peripheral irrelevant visual textures, whose position was systematically varied in the upper visual field (close, medium, or far relative to fixation). Either a neutral or a positive mood was reliably induced and maintained throughout the experimental session. The ERP results showed that the earliest retinotopic component following stimulus onset (C1) strongly varied in topography as a function of the position of the peripheral distractor, in agreement with a near-far spatial gradient. However, this effect was altered for participants in a positive relative to a neutral mood. On the contrary, positive mood did not modulate attention allocation for the central (task-relevant) stimuli, as reflected by the P300 component. We ran a control behavioral experiment confirming that positive emotion selectively impaired attention allocation to the peripheral distractors. These results suggest a mood-dependent tuning of position-specific encoding in V1 rapidly following stimulus onset. We discuss these results against the dominant broaden-and-build theory

    Does microbicide use in consumer products promote antimicrobial resistance? A critical review and recommendations for a cohesive approach to risk assessment

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    The increasing use of microbicides in consumer products is raising concerns related to enhanced microbicide resistance in bacteria and potential cross resistance to antibiotics. The recently published documents on this topic from the European Commission have spawned much interest to better understand the true extent of the putative links for the benefit of the manufacturers, regulators, and consumers alike. This white paper is based on a 2-day workshop (SEAC-Unilever, Bedford, United Kingdom; June 2012) in the fields of microbicide usage and resistance. It identifies gaps in our knowledge and also makes specific recommendations for harmonization of key terms and refinement/standardization of methods for testing microbicide resistance to better assess the impact and possible links with cross resistance to antibiotics. It also calls for a better cohesion in research in this field. Such information is crucial to developing any risk assessment framework on microbicide use notably in consumer products. The article also identifies key research questions where there are inadequate data, which, if addressed, could promote improved knowledge and understanding to assess any related risks for consumer and environmental safety

    Negative affective state mimics effects of perceptual load on spatial perception

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    Recent electrophysiological evidence has shown that perceptual load and negative affective state can produce very similar, early-attention gating effects in early visual areas, modulating the processing of peripheral stimuli. Here we assessed the question of whether or not these modulatory effects of perceptual load and negative affect (NA) lead to comparable changes in spatial perception abilities, which could be captured at the behavioral level. High perceptual load at fixation impaired the precise spatial localization of peripheral textures, relative to a low perceptual load condition. By contrast, the coarse spatial encoding of these peripheral stimuli was not load-dependent, under neutral affective conditions. The transient experience of NA was induced in an independent sample of participants, who showed decreased performance in the localization task, even at a low perceptual-load level. These results were observed in the absence of any systematic eye movement toward the peripheral textures. These findings suggest that spatial location perception is an attention-dependent, as well as state-dependent process, in the sense that NA, very much like load, can dynamically shape early spatial perceptual abilities. Although NA mimics load during spatial localization, we discuss the possibility that these two effects likely depend upon nonoverlapping brain networks
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