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

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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A. & Yablonka-Reuveni, Z. The skeletal muscle satellite cell: the stem cell that came in from the cold. J Histochem Cytochem 54, 1177–1191 (2006).McCord, M. C. & Aizenman, E. The role of intracellular zinc release in aging, oxidative stress, and Alzheimer’s disease. Front. Aging Neurosci. 6, 1–16 (2014).Dirksen, R. T. Sarcoplasmic reticulum–mitochondrial through-space coupling in skeletal muscle. This paper is one of a selection of papers published in this Special Issue, entitled 14th International Biochemistry of Exercise Conference – Muscles as Molecular and Metabolic. Appl. Physiol. Nutr. Metab. 34, 389–395 (2009).Groth, C., Sasamura, T., Khanna, M. R., Whitley, M. & Fortini, M. E. Protein trafficking abnormalities in Drosophila tissues with impaired activity of the ZIP7 zinc transporter Catsup. Development 140, 3018–3027 (2013).Ellis, C. D. et al. Zinc and the Msc2 zinc transporter protein are required for endoplasmic reticulum function. J. 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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

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

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)

Health risk as a consequence of exposure to trihalomethanes in swimming pool water

The article presents the formation and toxicity of trihalomethanes (THMs), substances belonging to Disinfection By- Products (DBP), formed as a result of pool water chlorination. Concentration of THMs in pool water has been standardised in Poland since recently. THMs concentration in pool water depends on multiple factors, mostly the method of water disinfection, including chlorine or organic substance (TOC) concentration. Apart from that, the level of exposure of swimmers to the toxic effect of THMs is affected by their content in the air of swimming pool halls, intensity of swimming as well as time of stay in the indoor swimming pool area. The water and air temperature as well as number of swimmers and time of their staying in pool have also significance influence on THMs concentration. In order to decrease the quantity of THMs in pool water, alternative disinfection agents are applied. Additionally, an efficiently operating pool ventilation system as well as proper water treatment method are also important. Because THMs are produced from organic material under the influence of the chlorination, the users can have an impact on decrease of the THMs concentration in pool water, for instance by taking a shower prior to entering the pool or by wearing a swim cap. The results of studies conducted in 2015 showed that the quantity of THMs in pool water depends on type of the pool (indoor, outdoor), water replacement frequency, water treatment system, chlorine dose and TOC content. In most collected samples, the THMs concentration exceeded the admissible norm (100 μg/L).W artykule omówiono powstawanie i działanie toksyczne trihalometanów (THM), związków należących do tzw. Disinfection By-Products (DBP), powstających w wyniku chlorowania wody w nieckach basenowych. Od niedawna w Polsce normuje się stężenie tych związków w wodzie basenowej. Stężenie THM w wodach w obiektach basenowych zależy od wielu czynników, głównie od sposobu dezynfekcji wody, w tym stężenia chloru oraz substancji organicznych (TOC). Poza tym na stopień narażenia pływaków na toksyczne działanie THM ma ich zawartość w powietrzu hali basenowych, intensywność pływania oraz czas przebywania na terenie pływalni. Ważna jest też temperatura wody i powietrza oraz ilość osób korzystających z basenu w danym czasie. W celu zmniejszenia ilości THM w wodzie basenowej stosuje się alternatywne środki dezynfekcji. Poza tym ważny jest sprawnie działający system wentylacji hali basenowej i odpowiedni sposób oczyszczania wody. Ponieważ THM są wytwarzane z udziałem substancji organicznych pod wpływem chlorowania, korzystający z basenów mogą wpłynąć na obniżenie poziomu THM, np. poprzez noszenie czepków oraz skorzystanie z prysznica przed wejściem do wody. Przeprowadzone w 2015 roku badania wykazały, że ilość THM w wodzie basenowej zależy od wielu czynników, w tym rodzaju basenu (kryty, odkryty), częstości wymian wody, rodzaju systemu jej oczyszczania, dawki chloru oraz zawartości TOC. W większości pobranych próbek stężenie THM przekraczało dopuszczalną normę (100 μg/L)

Fertilia: a European International Cooperation (INCO EU) project for monitoring trace elements in agricultural ecosystems

Background. A research project was financed starting in 1938 by the European Commission within the fourth Framework, involving International Cooperation (INCO EU) with Central European Countries. The project INCO Copernicus IC15-CT98-0124, acronym FERTILIA, addresses the relevance of trace metals for soil fertility. The participants come from four different countries: Italy, also acting as Coordinator, with the University of Parma, Department of Environmental Sciences, Division of Genetics and Environmental Biotechnology; Spain, with Consejo superior de Investigaciones Cientificas, Centre for the Biology of the Soil; Poland, with two partners, the Academy of Agriculture, Department of Microbiology and the University of Opole, Institute of Chemistry; finally, the Czech Republic, with the Academy of Sciences, Department of Environmental Analytical Chemistry. The participants bring together different expertise ranging from the analytical chemistry to soil microbiology up to plant genetic engineering and physic spectroscopy. The overall purpose of the three years project FERTILIA is to monitor the partitioning of trace elements in agricultural plants and vegetables grown on European soils subjected to different fertilisation regimes, taking into consideration all the biological, pedological and geochemical factors which can affect their form.Methods. Objectives and phases of the research are: determination of metal concentration in different sources of agricultural fertilisers (Action 1); physico-chemical analysis of metal concentration and aggregation in European soils (Action 2); analysis of metal content and partitioning ia different organisms taken from different levels of soil ecosystem (Action 3); morphological and functional effects of metals monitored with biomarkers microorganisms, invertebrates, and plants (Action 4); analysis of ecotoxicological and genotoxic effects of metals through the utilisation of model systems (Action 5); production and description of spatial models of soil ecosystems, by integrating geochemical and biological data within a Geographic Information System (GIS) (Action 6).Results. After the first year of research, Actions 1, 2, 3, and 4 reached a satisfactory set of experimental data. The last two actions are still in progress, because they require data from the first ones in order to be developed.Conclusions. Beside trying to show whether heavy metals can now in relevant amounts from fertilisers to soils and then to plants and soil microorganisms, this research project is trying to determine at which degree agricultural practices can modify soils metals content and the consequences this has on the crops quality