Real-time imaging of Leishmania mexicana-infected early phagosomes: a study using primary macrophages generated from green fluorescent protein-Rab5 transgenic mice

The small GTPase Rab5 is a key regulator of endosome/phagosome maturation and in intravesicular infections marks a phagosome stage at which decisions over pathogen replication or destruction are integrated. It is currently unclear whether Leishmania-infected phagosomes uniformly pass through a Rab5+ stage on their intracellular path to compartments with late endosomal/early lysosomal characteristics. Differences in routes and final compartments could have consequences for accessibility to antileishmanial drugs. Here, we generated a unique gfp-rab5 transgenic mouse model to visualize Rab5 recruitment to early parasite-containing phagosomes in primary host cells. Using real-time fluorescence imaging of phagosomes carrying Leishmania mexicana, we determined that parasite-infested phagosomes follow a uniform sequence of transient Rab5 recruitment. Residence in Rab5+ compartments was much shorter compared with phagosomes harboring latex beads. Furthermore, a comparative analysis of parasite life-cycle stages and mutants deficient in lpg1, the gene encoding the enzyme required for synthesis of the dominant surface lipophosphoglycan, indicated that parasite surface ligands and host cell receptors modulate pathogen residence times in Rab5+ phagosomes, but, as far as tested, had no significant effect on intracellular L. mexicana survival or replication.—Lippuner, C., Paape, D., Paterou, A., Brand, J., Richardson, M., Smith, A. J., Hoffmann, K., Brinkmann, V., Blackburn, C., Aebischer, T. Real-time imaging of Leishmania mexicana-infected early phagosomes: a study using primary macrophages generated from green fluorescent protein-Rab5 transgenic mice

Manipulating the Mouse Genome to Engineer Precise Functional Syntenic Replacements with Human Sequence

SummaryWe have devised a strategy (called recombinase-mediated genomic replacement, RMGR) to allow the replacement of large segments (>100 kb) of the mouse genome with the equivalent human syntenic region. The technique involves modifying a mouse ES cell chromosome and a human BAC by inserting heterotypic lox sites to flank the proposed exchange interval and then using Cre recombinase to achieve segmental exchange. We have demonstrated the feasibility of this approach by replacing the mouse α globin regulatory domain with the human syntenic region and generating homozygous mice that produce only human α globin chains. Furthermore, modified ES cells can be used iteratively for functional studies, and here, as an example, we have used RMGR to produce an accurate mouse model of human α thalassemia. RMGR has general applicability and will overcome limitations inherent in current transgenic technology when studying the expression of human genes and modeling human genetic diseases

Biochemical properties of Paracoccus denitrificans FnrP:Reactions with molecular oxygen and nitric oxide

In Paracoccus denitrificans, three CRP/FNR family regulatory proteins, NarR, NnrR and FnrP, control the switch between aerobic and anaerobic (denitrification) respiration. FnrP is a [4Fe-4S] cluster containing homologue of the archetypal O2 sensor FNR from E. coli and accordingly regulates genes encoding aerobic and anaerobic respiratory enzymes in response to O2, and also NO, availability. Here we show that FnrP undergoes O2-driven [4Fe-4S] to [2Fe-2S] cluster conversion that involves up to 2 O2 per cluster, with significant oxidation of released cluster sulfide to sulfane observed at higher O2 concentrations. The rate of the cluster reaction was found to be ~6-fold lower than that of E. coli FNR, suggesting that FnrP can remain transcriptionally active under microaerobic conditions. This is consistent with a role for FnrP in activating expression of the high O2 affinity cytochrome c oxidase under microaerobic conditions. Cluster conversion resulted in dissociation of the transcriptionally active FnrP dimer into monomers. Therefore, along with E. coli FNR, FnrP belongs to the subset of FNR proteins in which cluster type is correlated with association state. Interestingly, two key charged residues, Arg140 and Asp154, that have been shown to play key roles in the monomer-dimer equilibrium in E. coli FNR are not conserved in FnrP, indicating that different protomer interactions are important for this equilibrium. Finally, the FnrP [4Fe-4S] cluster is shown to undergo reaction with multiple NO molecules, resulting in iron nitrosyl species and dissociation into monomers

Information systems for collaborating versus transacting: Impact on manufacturing plant performance in the presence of demand volatility⋆

Research at the nexus of operations management and information systems suggests that manufacturing plants may benefit from the utilization of information systems for collaborating and transacting with suppliers and customers. The objective of this study is to examine the extent to which value generated by information systems for collaborating versus transacting is contingent upon demand volatility. We analyze a unique dataset assembled from non‐public U.S. Census Bureau data of manufacturing plants. Our findings suggest that when faced with volatile demand, plants employing information systems for collaborating with suppliers and customers experience positive and significant benefits to performance, in terms of both labor productivity and inventory turnover. In contrast, results suggest that plants employing information systems for transacting in volatile environments do not experience such benefits. Further exploratory analysis suggests that in the context of demand volatility, these two distinct dimensions of IT‐based integration have differing performance implications at different stages of the production process in terms of raw‐materials inventory and finished‐goods inventory, but not in terms of work‐in‐process inventory. Taken together, our study contributes to theoretical and managerial understanding of the contingent value of information systems in volatile demand conditions in the supply chain context.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/147128/1/joom313.pd

Spirit, mind and body: the archaeology of monastic healing

Archaeology and material culture are used in this chapter to consider how monastic experience responded to illness, ageing and disability. The approach taken is influenced by the material study of religion, which interrogates how bodies and things engage to construct the sensory experience of religion, and by practice-based approaches in archaeology, which examine the active role of space and material culture in shaping religious agency and embodiment. The archaeology of monastic healing focuses on the full spectrum of healing technologies, from managing the body in order to prevent illness, through to the treatment of the sick and preparation of the corpse for burial

Electroactivity across the cell wall of Gram-positive bacteria

Funding Information: This work was supported by national funds through FCT? Funda??o para a Ci?ncia e a Tecnologia, I.P. Project UIDB/04612/2020, UIDP/04612/2020 and PTDC/BIA-BQM/30176/2017, and by the European Union's Horizon 2020 research and innovation programme under grant agreement No 810856. Funding Information: This work was supported by national funds through FCT– Fundação para a Ciência e a Tecnologia, I.P., Project UIDB/04612/2020, UIDP/04612/2020 and PTDC/BIA-BQM/30176/2017, and by the European Union’s Horizon 2020 research and innovation programme under grant agreement No 810856. Publisher Copyright: © 2020 The Author(s) Copyright: Copyright 2021 Elsevier B.V., All rights reserved.The growing interest on sustainable biotechnological processes for the production of energy and industrial relevant organic compounds have increased the discovery of electroactive organisms (i.e. organisms that are able to exchange electrons with an electrode) and the characterization of their extracellular electron transfer mechanisms. While most of the knowledge on extracellular electron transfer processes came from studies on Gram-negative bacteria, less is known about the processes performed by Gram-positive bacteria. In contrast to Gram-negative bacteria, Gram-positive bacteria lack an outer-membrane and contain a thick cell wall, which were thought to prevent extracellular electron transfer. However, in the last decade, an increased number of Gram-positive bacteria have been found to perform extracellular electron transfer, and exchange electrons with an electrode. In this mini-review the current knowledge on the extracellular electron transfer processes performed by Gram-positive bacteria is introduced, emphasising their electroactive role in bioelectrochemical systems. Also, the existent information of the molecular processes by which these bacteria exchange electrons with an electrode is highlighted. This understanding is fundamental to advance the implementation of these organisms in sustainable biotechnological processes, either through modification of the systems or through genetic engineering, where the organisms can be optimized to become better catalysts.publishersversionpublishe

Can mindfulness and acceptance be learnt by self-help?: A systematic review and meta-analysis of mindfulness and acceptance-based self-help interventions

There is growing evidence that mindfulness and acceptance-based interventions have positive consequences for psychological and physical health. The most well-established of these interventions typically involve relatively large resource commitments, in terms of both the provider and participant. A number of recent studies have begun to explore whether the benefits of such interventions can be generalised to less intensive methods. Methods include pure and guided self-help utilising resources such as books and workbooks, computer programmes and applications and audio-visual materials. This paper presents a systematic review and meta-analysis of studies that have evaluated the effectiveness and acceptability of low-intensity interventions including mindfulness and acceptance-based components. Fifteen RCTs (7 ACT-based, 4 mindfulness-based and 4 multi-component interventions including elements of mindfulness and/or acceptance) were identified and reviewed. Interventions that included mindfulness and/or acceptance-based components produced significant benefits in comparison to control conditions on measures of mindfulness/acceptance, depression and anxiety with small to medium effect sizes. Engagement with the self-help interventions varied but on average two-thirds of participants completed post-intervention measures. Emerging research into low-intensity mindfulness and acceptance-based interventions is hopeful. Recommendations for research and practice are presente

Unstructured grid modelling of offshore wind farm impacts on seasonally stratified shelf seas

Shelf seas comprise approximately 7% of the world’s oceans and host enormous economic activity. Development of energy installations (e.g. Offshore Wind Farms (OWFs), tidal turbines) in response to increased demand for renewable energy requires a careful analysis of potential impacts. Recent remote sensing observations have identified kilometrescale impacts from OWFs. Existing modelling evaluating monopile impacts has fallen into two camps: small-scale models with individually resolved turbines looking at local effects; and large-scale analyses but with sub-grid scale turbine parameterisations. This work straddles both scales through a 3D unstructured grid model (FVCOM): wind turbine monopiles in the eastern Irish Sea are explicitly described in the grid whilst the overall grid domain covers the south-western UK shelf. Localised regions of decreased velocity extend up to 250 times the monopile diameter away from the monopile. Shelf-wide, the amplitude of the M2 tidal constituent increases by up to 7%. The turbines enhance localised vertical mixing which decreases seasonal stratification. The spatial extent of this extends well beyond the turbines into the surrounding seas. With significant expansion of OWFs on continental shelves, this work highlights the importance of how OWFs may impact coastal (e.g. increased flooding risk) and offshore (e.g. stratification and nutrient cycling) areas