Time- and dose dependent actions of cardiotonic steroids on transcriptome and intracellular content of Na+ and K+: a comparative analysis

Recent studies demonstrated that in addition to Na+,K+-ATPase inhibition cardiotonic steroids (CTSs) affect diverse intracellular signaling pathways. This study examines the relative impact of [Na+]i/[K+]i-mediated and -independent signaling in transcriptomic changes triggered by the endogenous CTSs ouabain and marinobufagenin (MBG) in human umbilical vein endothelial cells (HUVEC). We noted that prolongation of incubation increased the apparent affinity for ouabain estimated by the loss of [K+]i and gain of [Na+]i. Six hour exposure of HUVEC to 100 and 3,000 nM ouabain resulted in elevation of the [Na+]i/[K+]i ratio by ~15 and 80-fold and differential expression of 258 and 2185 transcripts, respectively. Neither [Na+]i/[K+]i ratio nor transcriptome were affected by 6-h incubation with 30 nM ouabain. The 96-h incubation with 3 nM ouabain or 30 nM MBG elevated the [Na+]i/[K+]i ratio by ~14 and 3-fold and led to differential expression of 880 and 484 transcripts, respectively. These parameters were not changed after 96-h incubation with 1 nM ouabain or 10 nM MBG. Thus, our results demonstrate that elevation of the [Na+]i/[K+]i ratio is an obligatory step for transcriptomic changes evoked by CTS in HUVEC. The molecular origin of upstream [Na+]i/[K+]i sensors involved in transcription regulation should be identified in forthcoming studies

Deoxygenation affects composition of membrane-bound proteins in human erythrocytes

Background/Aims: ATP release from erythrocyte plays a key role in hypoxia-induced elevation of blood flow in systematic circulation. We have previously shown that hemolysis contributes to erythrocyte ATP release triggered by several stimuli, including hypoxia, but the molecular mechanisms of hypoxia-increased membrane fragility remain unknown. Methods: In this study, we compared the action of hypoxia on hemolysis, ATP release and the composition of membrane-bound proteins in human erythrocytes. Results: Twenty minutes incubation of human erythrocytes in the oxygen-free environment increased the content of extracellular hemoglobin by ∼1.5 fold. Paired measurements of hemoglobin and ATP content in the same samples, showed a positive correlation between hemolysis and ATP release. Comparative analysis of SDS-PAGE electrophoresis of erythrocyte ghosts obtained under control and deoxygenated conditions revealed a ∼2-fold elevation of the content of membrane-bound protein with Mr of ∼60 kDa. Conclusion: Deoxygenation of human erythrocytes affects composition of membrane-bound proteins. Additional experiments should be performed to identify the molecular origin of 60 kDa protein and its role in the attenuation of erythrocyte integrity and ATP release in hypoxic conditions

Patterns and Temporal Dynamics of Natural Recombination in Noroviruses

Noroviruses infect a wide range of mammals and are the major cause of gastroenteritis in humans. Recombination at the junction of ORF1 encoding nonstructural proteins and ORF2 encoding major capsid protein VP1 is a well-known feature of noroviruses. Using all available complete norovirus sequences, we systematically analyzed patterns of natural recombination in the genus Norovirus both throughout the genome and across the genogroups. Recombination events between nonstructural (ORF1) and structural genomic regions (ORF2 and ORF3) were found in all analyzed genogroups of noroviruses, although recombination was most prominent between members of GII, the most common genogroup that infects humans. The half-life times of recombinant forms (clades without evidence of recombination) of human GI and GII noroviruses were 10.4 and 8.4–11.3 years, respectively. There was evidence of many recent recombination events, and most noroviruses that differed by more than 18% of nucleotide sequence were recombinant relative to each other. However, there were no distinct recombination events between viruses that differed by over 42% in ORF2/3, consistent with the absence of systematic recombination between different genogroups. The few inter-genogroup recombination events most likely occurred between ancient viruses before they diverged into contemporary genogroups. The recombination events within ORF1 or between ORF2/3 were generally rare. Thus, noroviruses routinely exchange full structural and nonstructural blocks of the genome, providing a modular evolution

Twisted Bilayer Graphene Quantum Dots for Chiral Nanophotonics

Twisted bilayer graphene (TBG) is a two-dimensional chiral material whose optical activity is remarkably strong for its atomic thickness. While the chiral optical properties of TBG are currently well understood, the optical activity of quantum dots (QDs) made of TBG has not been analyzed so far. Here, we comprehensively analyze the chiral optical properties of TBG QDs with hexagonal, triangular, and rhomboid symmetries. We show that the dissymmetry factors of TBG QDs can reach 10(-2), greatly exceeding the dissymmetry factors of semiconductor QDs and small chiral molecules. The strong optical activity of TBG QDs is due to the out-of-plane polarized transitions of the p(z) electrons and can be controlled by changing the size and shape of the QDs. These unique features make TBG QDs highly attractive for application in chiral nanophotonics

Microstructural and Hardness Behavior of H13 Tool Steel Manufactured by Ultrasound-Assisted Laser-Directed Energy Deposition

Metal additive manufacturing (AM) by Laser-Directed Energy Deposition (L-DED) usually results in the formation of textured columnar grains along the build direction, leading to anisotropic mechanical properties. This can negatively affect the intended application of the product. Anisotropy can be eliminated by modifying the material through an additional exposure to ultrasound (US-assisted) during the L-DED process. In this paper, a multi-track sample was manufactured from AISI H13 (TLS Technik, Bitterfeld-Wolfen, Germany) tool steel by a US-assisted (28 kHz) L-DED process using a specially designed cooling system. The study also included post-process annealing and quenching with the tempering heat treatment of the modified steel, resulting in the retention of the properties, as confirmed by hardness measurements. XRD analysis was used to measure the structural parameters of the unit cell, and the hardness properties were measured in two directions: longitudinally and parallel to the deposition direction. It was found that US-assisted L-DED allows us to obtain a more isotropic structure with an equal size of the coherent scattering region in two printing directions, and to reduce the residual stresses in the material. The anisotropy of the hardness was significantly reduced, with 636 and 640 HV found between the XY and XZ planes. Based on the obtained hardness data, it should be noted that some of the heat treatments studied herein can also result in a decrease in the anisotropy of the properties, similarly to the US-assisted effect.Team Vera Popovic

Laser Ablation Synthesis and Characterization of Tb2O3 Nanoparticles for Magneto-Optical Ceramics

In this study, nano-sized individual Tb2O3 particles synthesized by the laser ablation method were extensively characterized and assessed as suitable precursors for the fabrication of transparent magneto-optical ceramics without requiring the introduction of grain growth inhibitors and stabilizing additives. The as-produced powder comprised 13 nm particles with a spherical shape and monoclinic crystal structure, whose full transformation into cubic phase was achieved after heating at 950 °C and 700 °C under vacuum and Ar gas, respectively. After subjecting the nanopowder compact to pre-sintering at 1350 °C, the microstructural features were investigated along with their correlation to the optical transmittance of Tb2O3 ceramic hot isostatically pressed (HIPed) for 2 h at 1450 °C under 200 MPa. The as-HIPed sample had a brownish color, with an optical transmittance of 65.3% at a wavelength of 1060 nm and an average grain size of 14 μm. The Verdet constant measured at wavelengths of 633 nm and 1060 nm was 471 rad T−1 m−1 and 142 rad T−1 m−1 to confirm a very high content of magneto-active Tb3+ ions. The obtained results indicate that laser ablation synthesis of nanoparticles followed by pre-sintering and HIP is a promising approach for the manufacture of magneto-optical Tb2O3 ceramics without specific sintering aids

Laser Ablation Synthesis and Characterization of Tb₂O₃ Nanoparticles for Magneto-Optical Ceramics

In this study, nano-sized individual Tb2O3 particles synthesized by the laser ablation method were extensively characterized and assessed as suitable precursors for the fabrication of transparent magneto-optical ceramics without requiring the introduction of grain growth inhibitors and stabilizing additives. The as-produced powder comprised 13 nm particles with a spherical shape and monoclinic crystal structure, whose full transformation into cubic phase was achieved after heating at 950 °C and 700 °C under vacuum and Ar gas, respectively. After subjecting the nanopowder compact to pre-sintering at 1350 °C, the microstructural features were investigated along with their correlation to the optical transmittance of Tb2O3 ceramic hot isostatically pressed (HIPed) for 2 h at 1450 °C under 200 MPa. The as-HIPed sample had a brownish color, with an optical transmittance of 65.3% at a wavelength of 1060 nm and an average grain size of 14 μm. The Verdet constant measured at wavelengths of 633 nm and 1060 nm was 471 rad T−1 m−1 and 142 rad T−1 m−1 to confirm a very high content of magneto-active Tb3+ ions. The obtained results indicate that laser ablation synthesis of nanoparticles followed by pre-sintering and HIP is a promising approach for the manufacture of magneto-optical Tb2O3 ceramics without specific sintering aids

Time- and dose dependent actions of cardiotonic steroids on transcriptome and intracellular content of Na+ and K+: a comparative analysis

Recent studies demonstrated that in addition to Na+,K+-ATPase inhibition cardiotonic steroids (CTSs) affect diverse intracellular signaling pathways. This study examines the relative impact of [Na+]i/[K+]i-mediated and -independent signaling in transcriptomic changes triggered by the endogenous CTSs ouabain and marinobufagenin (MBG) in human umbilical vein endothelial cells (HUVEC). We noted that prolongation of incubation increased the apparent affinity for ouabain estimated by the loss of [K+]i and gain of [Na+]i. Six hour exposure of HUVEC to 100 and 3,000 nM ouabain resulted in elevation of the [Na+]i/[K+]i ratio by ~15 and 80-fold and differential expression of 258 and 2185 transcripts, respectively. Neither [Na+]i/[K+]i ratio nor transcriptome were affected by 6-h incubation with 30 nM ouabain. The 96-h incubation with 3 nM ouabain or 30 nM MBG elevated the [Na+]i/[K+]i ratio by ~14 and 3-fold and led to differential expression of 880 and 484 transcripts, respectively. These parameters were not changed after 96-h incubation with 1 nM ouabain or 10 nM MBG. Thus, our results demonstrate that elevation of the [Na+]i/[K+]i ratio is an obligatory step for transcriptomic changes evoked by CTS in HUVEC. The molecular origin of upstream [Na+]i/[K+]i sensors involved in transcription regulation should be identified in forthcoming studies

Deoxygenation affects composition of membrane-bound proteins in human erythrocytes

Background/Aims: ATP release from erythrocyte plays a key role in hypoxia-induced elevation of blood flow in systematic circulation. We have previously shown that hemolysis contributes to erythrocyte ATP release triggered by several stimuli, including hypoxia, but the molecular mechanisms of hypoxia-increased membrane fragility remain unknown. Methods: In this study, we compared the action of hypoxia on hemolysis, ATP release and the composition of membrane-bound proteins in human erythrocytes. Results: Twenty minutes incubation of human erythrocytes in the oxygen-free environment increased the content of extracellular hemoglobin by ∼1.5 fold. Paired measurements of hemoglobin and ATP content in the same samples, showed a positive correlation between hemolysis and ATP release. Comparative analysis of SDS-PAGE electrophoresis of erythrocyte ghosts obtained under control and deoxygenated conditions revealed a ∼2-fold elevation of the content of membrane-bound protein with Mr of ∼60 kDa. Conclusion: Deoxygenation of human erythrocytes affects composition of membrane-bound proteins. Additional experiments should be performed to identify the molecular origin of 60 kDa protein and its role in the attenuation of erythrocyte integrity and ATP release in hypoxic conditions