Calcium signaling is essential for the differentiation of many cell types, including skeletal muscle cells, but its mechanisms remain elusive. Here we demonstrate a crucial role for nicotinic acid adenine dinucleotide phosphate (NAADP) signaling in skeletal muscle differentiation. Although the inositol trisphosphate pathway may have a partial role to play in this process, the ryanodine signaling cascade is not involved. In both skeletal muscle precursors and C2C12, cells interfering with NAADP signaling prevented differentiation, whereas promoting NAADP signaling potentiated differentiation. Moreover, siRNA knockdown of two-pore channels, the target of NAADP, attenuated differentiation. The data presented here strongly suggest that in myoblasts, NAADP acts at acidic organelles on the recently discovered two-pore channels to promote differentiation.

In recent years, many new opioids, particularly fentanyl analogues, have appeared on the drug market. The extreme potency of even low doses of these compounds leads to numerous fatal poisonings. This also results in the fact that only sophisticated techniques are capable of detecting fentanyl analogues at concentrations that can be expected in blood. In this context, the purpose of this study was to develop a fast liquid chromatography‐tandem mass spectrometry screening method for the detection of fentanyl analogues, and other new synthetic opioid receptor agonists in whole blood. Blood samples were extracted with ethyl acetate under basic conditions. The separation was achieved with the gradient of the mobile phase composition and the gradient of the flow rate in 13 minutes. The detection of all compounds was based on dynamic multiple reaction monitoring. Most of the compounds were well differentiated by their retention times and/or transitions; however, separation of some isomers has not been achieved. The validation was performed for 21 compounds. The limits of detection were in the range 0.01‐0.20 ng/mL. The developed procedure enables simultaneous qualitative screening, detection and identification of 38 fentanyl analogues and five other new opioids. The method was implemented to analyze authentic samples (positive; n = 3) demonstrating its suitability for this application. The procedure can be easily expanded to include new emerging opioids, which is an indispensable advantage in the dynamically developing drug market. The developed protocol can be adopted for routine work in both forensic and clinical analytical laboratories worldwide.

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The aim of this study was to evaluate the effect of low-energy laser irradiation, coenzyme Q10 and vitamin E supplementation on the apoptosis of macrophages and muscle precursor cells during skeletal muscle regeneration after bupivacaine-induced injury. The experiment was conducted on 75 gilts, divided into 5 experimental groups: I -control, II -low-energy laser irradiation, III -coenzyme Q10, IV -coenzyme Q10 and vitamin E, V -vitamin E. Muscle necrosis was induced by injection of 0.5% bupivacaine hydrochloride. The animals were euthanized on subsequent days after injury. Samples were formalin fixed and processed routinely for histopathology. Apoptosis was detected using the TUNEL method. The obtained results indicate that low-energy laser irradiation has a beneficial effect on macrophages and muscle precursor cell activity during muscle post-injury regeneration and protects these cells against apoptosis. Vitamin E has a slightly lower protective effect, limited mainly to the macrophages. Coenzyme Q10 co-supplemented with vitamin E increases the activity of macrophages and muscle precursor cells, myotube and young muscle formation. Importantly, muscle precursor cells seem to be more sensitive to apoptosis than macrophages in the environment of regenerating damaged muscle.

During the last decade, tree species mixing has been widely supported as a silvicultural approach to reduce drought stress. However, little is known on the influence of tree species mixing on physical properties and the water storage capacity of forest soils (including the forest floor). Thus, the study aimed to analyze the effect of mixing pine needles and oak leaves and mixing fir needles and beech leaves on hydro-physical properties of the litter layer during laboratory tests. We used fir-beech and pine-oak litter containing various shares of conifer needles (i.e., 0, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90 and 100%) to determine the influence of the needle admixture on bulk density, total porosity, macroporosity, water storage capacity, the amount of water stored in pores between organic debris and the degree of saturation of mixed litter compared to broadleaf litter (oak or beech). We found that the admixture of fir needles increased the bulk density of litter from 7.9% with a 5% share of needles to 55.5% with a 50% share (compared to pure beech litter), while the share of pine needles < 40% caused a decrease in bulk density by an average of 3.0–11.0% (compared to pure oak litter). Pine needles decreased the water storage capacity of litter by about 13–14% with the share of needles up to 10% and on average by 28% with the 40 and 50% shares of pine needles in the litter layer. Both conifer admixtures reduced the amount of water stored in the pores between organic debris (pine needles more than fir needles).