186 research outputs found
Calcium signals and calpain-dependent necrosis are essential for release of coxsackievirus B from polarized intestinal epithelial cells
Coxsackievirus B (CVB), a member of the enterovirus family, targets the polarized epithelial cells lining the intestinal tract early in infection. Although the polarized epithelium functions as a protective barrier, this barrier is likely exploited by CVB to promote viral entry and subsequent egress. Here we show that, in contrast to nonpolarized cells, CVB-infected polarized intestinal Caco-2 cells undergo nonapoptotic necrotic cell death triggered by inositol 1,4,5-trisphosphate receptor-dependent calcium release. We further show that CVB-induced cellular necrosis depends on the Ca 2+-activated protease calpain-2 and that this protease is involved in CVB-induced disruption of the junctional complex and rearrangements of the actin cytoskeleton. Our study illustrates the cell signaling pathways hijacked by CVB, and perhaps other viral pathogens, to promote their replication and spread in polarized cell types. © 2011 Bozym et al.published_or_final_versio
The study of heavy metals leaching from waste foundry sands using a one-step extraction
There are a number of leaching test which are used to evaluate the effect of foundry waste disposal on the environment (TCLP, SPLP, ASTM at al.). Because the spent foundry sand are usually deposited at the ground level and they have a similar structure to the soil, survey mobility of metals using the same methods seems appropriate. One-step extraction allows for the evaluation of the mobility and bioavailability of metals in soil and waste. Waste foundry sands have been successfully used as a component in manufactured soils in U.S., but concern over metal contamination must be eliminated before considering this direction of use. The study evaluated the leaching of heavy metals (Cd, Pb, Cu, Zn, Cr, Ni) from deposited waste foundry sands. The overall, as well as heavy metals were extracted by different type of extractants: H2O, CH3COOH, HCl, EDTA, MgCl2 and NaCOOH. These extractants are most commonly used to study the mobility and bioavailability of metals in soil and waste. In the present study applicable standards and methodology described in the literature in analysis were used. The results allowed to evaluate the bioavailability of metals leached from those wastes
HDAC8 substrates: Histones and beyond
The lysine deacetylase family of enzymes (HDACs) was first demonstrated to catalyze deacetylation of acetyllysine residues on histones. In subsequent years, HDACs have been shown to recognize a large pool of acetylated nonhistone proteins as substrates. Recently, thousands of acetylated proteins have been discovered, yet in most cases, the HDAC that catalyzes deacetylation in vivo has not been identified. This gap has created the need for better in vivo, in vitro, and in silico approaches for determining HDAC substrates. While HDAC8 is the best kinetically and structurally characterized HDAC, few efficient substrates have yet been substantiated in vivo. In this review, we delineate factors that may be important for determining HDAC8 substrate recognition and catalytic activity, including structure, complex formation, and post‐translational modifications. This summary provides insight into the challenges of identifying in vivo substrates for HDAC8, and provides a good vantage point for understanding the variables important for predicting HDAC substrate recognition. © 2012 Wiley Periodicals, Inc. Biopolymers 99: 112–126, 2013.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/94512/1/22135_ftp.pd
Zinc uptake promotes myoblast differentiation via Zip7 transporter and activation of Akt signalling transduction pathway
[EN] Myogenic regeneration occurs through a chain of events beginning with the output of satellite cells from quiescent state, formation of competent myoblasts and later fusion and differentiation into myofibres. Traditionally, growth factors are used to stimulate muscle regeneration but this involves serious off-target effects, including alterations in cell homeostasis and cancer. In this work, we have studied the use of zinc to trigger myogenic differentiation. We show that zinc promotes myoblast proliferation, differentiation and maturation of myofibres. We demonstrate that this process occurs through the PI3K/Akt pathway, via zinc stimulation of transporter Zip7. Depletion of zinc transporter Zip7 by RNA interference shows reduction of both PI3K/Akt signalling and a significant reduction of multinucleated myofibres and myotubes development. Moreover, we show that mature myofibres, obtained through stimulation with high concentrations of zinc, accumulate zinc and so we hypothesise their function as zinc reservoirs into the cell.P.R. and R.S. acknowledges support from the Spanish Ministry of Economy and Competitiveness (MINECO) (MAT2015-69315-C3-1-R). P.R. acknowledges the Fondo Europeo de Desarrollo Regional (FEDER). CIBER-BBN is an initiative funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. R.S. acknowledges the support from the Spanish MECD through the PRX16/00208 grant. MSS acknowledges support from the European Research Council (ERC - HealInSynergy 306990) and the UK Engineering and Physical Sciences Research Council (EPSRC - EP/P001114/1)Mnatsakanyan, H.; Sabater I Serra, R.; Rico Tortosa, PM.; Salmerón Sánchez, M. (2018). 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Release of Intracellular Calcium Stores Facilitates Coxsackievirus Entry into Polarized Endothelial Cells
Group B coxsackieviruses (CVB) are associated with viral-induced heart disease and are among the leading causes of aseptic meningitis worldwide. Here we show that CVB entry into polarized brain microvasculature and aortic endothelial cells triggers a depletion of intracellular calcium stores initiated through viral attachment to the apical attachment factor decay-accelerating factor. Calcium release was dependent upon a signaling cascade that required the activity of the Src family of tyrosine kinases, phospholipase C, and the inositol 1,4,5-trisphosphate receptor isoform 3. CVB-mediated calcium release was required for the activation of calpain-2, a calcium-dependent cysteine protease, which controlled the vesicular trafficking of internalized CVB particles. These data point to a specific role for calcium signaling in CVB entry into polarized endothelial monolayers and highlight the unique signaling mechanisms used by these viruses to cross endothelial barriers
Disulfiram-induced cytotoxicity and endo-lysosomal sequestration of zinc in breast cancer cells
Disulfiram, a clinically used alcohol-deterrent has gained prominence as a potential anti-cancer agent due to its impact on copper-dependent processes. Few studies have investigated zinc effects on disulfiram action, despite it having high affinity for this metal. Here we studied the cytotoxic effects of disulfiram in breast cancer cells, and its relationship with both intra and extracellular zinc. MCF-7 and BT474 cancer cell lines gave a striking time-dependent biphasic cytotoxic response between 0.01 and 10 μM disulfiram. Co-incubation of disulfiram with low-level zinc removed this effect, suggesting that availability of extracellular zinc significantly influences disulfiram efficacy. Live-cell confocal microscopy using fluorescent endocytic probes and the zinc dye Fluozin-3 revealed that disulfiram selectively and rapidly increased zinc levels in endo-lysosomes. Disulfiram also caused spatial disorganization of late endosomes and lysosomes, suggesting they are novel targets for this drug. This relationship between disulfiram toxicity and ionophore activity was consolidated via synthesis of a new disulfiram analog and overall we demonstrate a novel mechanism of disulfiram-cytotoxicity with significant clinical implications for future use as a cancer therapeutic
Imaging the boundaries—innovative tools for microscopy of living cells and real-time imaging
Recently, light microscopy moved back into the spotlight, which is mainly due to the development of revolutionary technologies for imaging real-time events in living cells. It is truly fascinating to see enzymes “at work” and optically acquired images certainly help us to understand biological processes better than any abstract measurements. This review aims to point out elegant examples of recent cell-biological imaging applications that have been developed with a chemical approach. The discussed technologies include nanoscale fluorescence microscopy, imaging of model membranes, automated high-throughput microscopy control and analysis, and fluorescent probes with a special focus on visualizing enzyme activity, free radicals, and protein–protein interaction designed for use in living cells
Zmiany parametrow fizykochemicznych osadu sciekowego w czasie wermikompostowania
Results of assessment of raw sewage sludge, vermicompost and earthworms from Waste Water Treatment Plant in Kluczbork are presented in this paper. Vermicomposting process improved sewage sludge from greasy to soily structure. It affected the level of organic matter, total N, Na, Cd, Zn during the process. The Cr and Cd contents remained at the same level. However, the increase of the level of dry matter, and the content of P, K, Ca, Mg, Pb, Cu was observed. Heavy metals cumulating in earthworms depended on the type of metal and their content in some cases (Cd, Pb, Ni) was higher than in vermicompost.W pracy przedstawiono wyniki badań osadów ściekowych i wermikompostu oraz dżdżownic pochodzących z Miejskiej Oczyszczalni Ścieków w Kluczborku. Proces wermikompostwania poprawił strukturę osadu z mazistej na ziemistą. Podczas procesu zauważono zmniejszenie się zawartości substancji organicznej, N ogólnego, Na, Cd, Zn. Zawartość Cr i Ni utrzymywała się na podobnym poziomie. Natomiast wzrosła zawartość suchej masy (zmniejszenie uwodnienia i objętości), P, K, Ca, Mg, Pb, Cu. Kumulacja metali ciężkich w dżdżownicach zależała od rodzaju metalu i w kilku przypadkach przekraczała zawartość w osadzie (Cd, Pb, Ni)
The Functionalized Graphene-Ionic Liquid Interface for Electrochemical Double-Layer Capacitors
Functionalized graphene sheets (FGSs) are promising electrode materials for energy-dense electrochemical double-layer capacitors (EDLCs) as they can exhibit high electrical conductivity, specific surface area and intrinsic capacitance. However, engineering electrodes that capitalize on these attributes is challenging as FGSs tend to re-stack during processing, reducing the surface area available for capacitive energy storage. Furthermore, though room temperature ionic liquids (RTILs) have been introduced as next-generation electrolytes for EDLCs, a fundamental understanding of the capacitance of the FGS-electrolyte interface for concentrated electrolytes is lacking. This dissertation aims to develop approaches to produce energy-dense, FGS-based EDLC electrodes and elucidate the fundamental behavior of the electrochemical double-layer of the FGS-electrolyte interface.
A simple ¿bottom-up¿ approach was developed to evaporatively consolidate mixtures of graphene oxide, alcohols, and RTILs, producing thin-film electrodes having an energy density as high as 6.3 Wh/kg at 3.0 V, based on the total mass of the electrode. To improve the energy density, dehydrated sucrose nanoparticles (0.5-2.5 nm) were added to serve as inter-sheet spacers. Liquid-cell atomic force microscopy revealed that these nanoparticles adsorb onto graphene oxide in an aqueous environment, providing a physical barrier to inter-sheet collapse due to van der Waals and capillary forces during solvent evaporation. Thin film electrodes were prepared having a gravimetric capacitance as high as 330 F/gFGS and, by tuning the nanoparticle and RTIL contents, an energy density of 13.3 Wh/kg was achieved. This energy density is the highest reported to date for an FGS-based electrode with a solid spacer.
Lastly, electrochemical impedance spectroscopy was used to measure the effect of diluting an RTIL with miscible organic solvents on the intrinsic capacitance of the glassy carbon-electrolyte interface. Diluting 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide with organic solvents revealed a maximum in the minimum differential capacitance near 5-10 mol% RTIL that was not predicted by classical electrochemical double-layer theory. Additionally, diluting with a low-dielectric constant solvent, 1,2-dichloroethane, resulted in the largest increases in capacitance near the open circuit potential, compared to acetonitrile and propylene carbonate. These results provide insight into the electrochemical double-layer for concentrated electrolytes and reveal the potential for the enhancement of double-layer capacitance through dilution
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