Microtubule rescue control by drugs and MAPs examined with in vitro pedestal assay.

Microtubules are essential cytoskeletal polymers, which exhibit stochastic transitions between assembly and disassembly, known as catastrophes and rescues. Understanding of catastrophes, rescues, and their control by drugs and microtubule associated proteins (MAPs) has been informed by in vitro reconstitutions of microtubule dynamics. In such experiments microtubules are typically observed on a flat surface of the coverslip. In contrast, we have recently proposed a modified setup in which microtubules assemble from stabilized seeds, overhanging from microfabricated pedestals, so that their dynamic extensions are fully isolated from contact with the coverslip. This assay allows to eliminate potential artifacts, which may substantially affect the frequency of microtubule rescues in vitro. Here we use the pedestal assay to study the sensitivity of microtubules to paclitaxel, one of the best-known inhibitors of microtubule dynamics. By comparing observations in the conventional and the pedestal assays, we find that microtubule dynamics are substantially more sensitive to paclitaxel when the polymers can contact the coverslip. We interpret this as a consequence of the coverslip-induced microtubule assembly perturbation, leading to formation of lattice with defects, and thereby enhancing the efficiency of paclitaxel binding to microtubules in the conventional assay. To test this idea, we use vinblastine, another small-molecule inhibitor, which had been previously shown to cause microtubule growth perturbations. We find that in the pedestal assay vinblastine sensitizes microtubules to paclitaxel to the level, observed in the conventional assay. Interestingly, a minimal fragment of MAP called CLASP2, a previously characterized rescue factor, has a strong effect on microtubule rescues, regardless of the type of assay. Overall, our study underscores the role of microtubule damage in promoting rescues and highlights the utility of the in vitro pedestal assay to study microtubule dynamics modulation by tubulin inhibitors and MAPs.

Monaural and dichotic forward masking in the dolphin's auditory system.

Short-latency auditory-evoked potentials (AEPs) were recorded non-invasively in the bottlenose dolphin Tursiops truncatus. The stimuli were two sound clicks that were played either monaurally (both clicks to one and the same acoustic window) or dichotically (the leading stimulus (masker) to one acoustic window and the delayed stimulus (test) to the other window). The ratio of the levels of the two stimuli was 0, 10, or 20 dB (at 10 and 20 dB, the leading stimulus was of a higher level). The inter-stimulus intervals (ISIs) varied from 0.15 to 10 ms. The test response magnitude was assessed by correlation analysis as a percentage of the control (non-masked) response. At monaural stimulation, the test response was of a constant magnitude (5-6% of the control) at ISIs of 0.15-0.3 ms and recovered at longer ISIs. At dichotic stimulation, the deepest suppression of the test response occurred at ISIs of 0.5-0.7 ms. The response was slightly suppressed at short ISIs (0.15-0.3 ms) and recovered at ISIs longer than 0.5-0.7 ms. The relation of parameters of the forward masking to echolocation in dolphins is discussed.

Glass Film Formation on GOES Surface during High-Temperature Annealing: The Mechanism with Amorphous Phase Formation.

Ceramic insulation coating (glass film) is an important constituent of grain-oriented electrical steel (GOES) designed for use in transformers. Within the scope of this study, the glass film was obtained by means of interaction between the surface of GOES containing 0.5 wt. % Cu and a heat-resistant MgO coating during annealing up to 1100 °C in the 75%H2 + 25%N2 atmosphere. The structure of glass film was analyzed using X-ray diffraction, glow-discharge optical emission spectroscopy, scanning probe microscopy, scanning electron microscopy, differential scanning calorimetry and thermodynamic calculations. After annealing, the glass film contained the following phases: crystalline (MgFe)2SiO4 and amorphous Fe-based solid solutions. The multi-stage mechanism of the glass film formation on GOES surface during high-temperature annealing was determined.

Theory of tip structure-dependent microtubule catastrophes and damage-induced microtubule rescues.

Microtubules are essential cytoskeletal polymers that exhibit stochastic switches between tubulin assembly and disassembly. Here, we examine possible mechanisms for these switches, called catastrophes and rescues. We formulate a four-state Monte Carlo model, explicitly considering two biochemical and two conformational states of tubulin, based on a recently conceived view of microtubule assembly with flared ends. The model predicts that high activation energy barriers for lateral tubulin interactions can cause lagging of curled protofilaments, leading to a ragged appearance of the growing tip. Changes in the extent of tip raggedness explain some important but poorly understood features of microtubule catastrophe: weak dependence on tubulin concentration and an increase in its probability over time, known as aging. The model predicts a vanishingly rare frequency of spontaneous rescue unless patches of guanosine triphosphate tubulin are artificially embedded into microtubule lattice. To test our model, we used in vitro reconstitution, designed to minimize artifacts induced by microtubule interaction with nearby surfaces. Microtubules were assembled from seeds overhanging from microfabricated pedestals and thus well separated from the coverslip. This geometry reduced the rescue frequency and the incorporation of tubulins into the microtubule shaft compared with the conventional assay, producing data consistent with the model. Moreover, the rescue positions of microtubules nucleated from coverslip-immobilized seeds displayed a nonexponential distribution, confirming that coverslips can affect microtubule dynamics. Overall, our study establishes a unified theory accounting for microtubule assembly with flared ends, a tip structure-dependent catastrophe frequency, and a microtubule rescue frequency dependent on lattice damage and repair.

Forward masking in a bottlenose dolphin Tursiops truncatus: dependence on azimuthal positions of the masker and test sources.

Forward masking was investigated by the auditory evoked potentials (AEP) method in a bottlenose dolphin Tursiops truncatus using stimulation by two successive acoustic pulses (the masker and test) projected from spatially separated sources. The positions of the two sound sources either coincided with or were symmetrical relative to the head axis at azimuths from 0 to ± 90°. AEPs were recorded either from the vertex or from the lateral head surface next to the auditory meatus. In the last case, the test source was ipsilateral to the recording side, whereas the masker source was either ipsi- or contralateral. For lateral recording, AEP release from masking (recovery) was slower for the ipsi- than for the contralateral masker source position. For vertex recording, AEP recovery was equal both for the coinciding positions of the masker and test sources and for their symmetrical positions relative to the head axis. The data indicate that at higher levels of the auditory system of the dolphin, binaural convergence makes the forward masking nearly equal for ipsi- and contralateral positions of the masker and test.

Industry 4.0 and Digitalisation in Healthcare.

Industry 4.0 in healthcare involves use of a wide range of modern technologies including digitisation, artificial intelligence, user response data (ergonomics), human psychology, the Internet of Things, machine learning, big data mining, and augmented reality to name a few. The healthcare industry is undergoing a paradigm shift thanks to Industry 4.0, which provides better user comfort through proactive intervention in early detection and treatment of various diseases. The sector is now ready to make its next move towards Industry 5.0, but certain aspects that motivated this review paper need further consideration. As a fruitful outcome of this review, we surveyed modern trends in this arena of research and summarised the intricacies of new features to guide and prepare the sector for an Industry 5.0-ready healthcare system.

Microstructural Features in Multicore Cu-Nb Composites.

The study is devoted to heavily drawn multicore Cu-18Nb composites of cylindrical and rectangular shapes. The composites were fabricated by the melt-and-deform method, namely, 600 in situ rods of Cu-18%Nb alloy were assembled in a copper shell and cold-drawn to a diameter of 15.4 mm (e = 10.2) and then rolled into a rectangular shape the size of 3 × 5.8 mm (e = 12.5). The specimens were analyzed from the viewpoints of their microstructure, microhardness, and thermal stability. The methods of SEM, TEM, X-ray analysis, and microhardness measurements were applied. It is demonstrated that, at higher strain, the fiber texture 110Nb∥ 111Cu∥ DD (drawing direction), characteristic of this material, becomes sharper. The distortions of niobium lattice can be observed, namely, the 110 Nb interplanar distance is broadened in longitudinal direction of specimens and compacted in transverse sections. The copper matrix lattice is distorted as well, though its distortions are much less pronounced due to its recrystallization. Evolution of microstructure under annealing consists mainly in the coagulation of ribbon-like Nb filaments and in the vanishing of lattice distortions. The structural changes in Nb filaments start at 300-400 °C, then develop actively at 600 °C and cause considerable decrease of strength at 700-800 °C.

The efficacy and safety of levilimab in severely ill COVID-19 patients not requiring mechanical ventilation: results of a multicenter randomized double-blind placebo-controlled phase III CORONA clinical study.

OBJECTIVE AND DESIGN: The aim of this double-blind, placebo-controlled, phase III CORONA clinical trial was to evaluate the efficacy and safety of IL-6 receptor inhibitor levilimab (LVL) in subjects with severe COVID-19. SUBJECTS: The study included 217 patients. The eligible were men and non-pregnant women aged 18 years or older, hospitalized for severe COVID-19 pneumonia. TREATMENT: 206 subjects were randomized (1:1) to receive single subcutaneous administration of LVL 324 mg or placebo, both in combination with standard of care (SOC). 204 patients received allocated therapy. After the LVL/placebo administration in case of deterioration of symptoms, the investigator could perform a single open-label LVL 324 mg administration as the rescue therapy. METHODS: The primary efficacy endpoint was the proportion of patients with sustained clinical improvement on the 7-category ordinal scale on Day 14. All efficacy data obtained after rescue therapy administration were considered missing. For primary efficacy analysis, all subjects with missing data were considered non-responders. RESULTS: 63.1% and 42.7% of patients in the LVL and in the placebo groups, respectively, achieved sustained clinical improvement on Day 14 (P = .0017). The frequency of adverse drug reactions was comparable between the groups. CONCLUSION: In patients with radiologically confirmed SARS-CoV-2 pneumonia, requiring or not oxygen therapy (but not ventilation) with no signs of other active infection administration of LVL + SOC results in an increase of sustained clinical improvement rate. TRAIL REGISTRATION: The trial is registered at the US National Institutes of Health (ClinicalTrials.gov; NCT04397562).

Texturing and Phase Evolution in Ti-6Al-4V: Effect of Electron Beam Melting Process, Powder Re-Using, and HIP Treatment.

The research demonstrates microstructural changes and development of specific texture in Ti-6Al-4V specimens produced by electron beam melting (EBM) under different conditions. The effect of two factors, namely, raw material (powder) recycling and hot isostatic pressing (HIP), on the EBM produced samples structure and properties, has been explored. The as-printed and treated samples were investigated using electron backscattered diffraction (EBSD) analysis. Modification of mechanical properties after the EBM and HIP are explained by the EBSD data on microstructural phenomena and phase transformations. The work is devoted to assessing the possibility of reusing the residual titanium alloy powder for the manufacture of titanium components by the combination of EBM and HIP methods.

Corrosion Resistance of Al-CNT Metal Matrix Composites.

The design of aluminium-graphite metal matrix composites (MMCs) with advanced mechanical properties and high corrosion resistance is in demand for aerospace, transportation, and industrial applications. Breakthroughs in this field are limited due to the tendency of aluminium-graphite MMCs to corrode. In the present research, aluminium-based MMCs were produced by a relatively novel combined two-staged method. Multiwall carbon nanotubes (MWCNTs) were added into molten Al1070 and processed by high-pressure die casting followed by cyclic extrusion. For the composites produced by this method, it was previously demonstrated that mechanical properties are improved in comparison with pure aluminium alloys. In the current study, the manufactured Al-MWCNT composites were investigated by electrochemical tests (such as open circuit potential), potentiodynamic tests, linear polarization tests, and electrochemical impedance spectra to understand the corrosion resistance of the obtained composite material. The experimental testing of the corrosion resistance of Al-MWCNT MMCs showed that due to the advantages of the fabrication method, the addition of CNTs to aluminium does not cause a radical decrease of corrosion resistance.

Complex Concentrated Alloys for Substitution of Critical Raw Materials in Applications for Extreme Conditions.

The paper is proposing a mini-review on the capability of the new complex concentrated alloys (CCAs) to substitute or reduce the use of critical raw materials in applications for extreme conditions. Aspects regarding the regulations and expectations formulated by the European Union in the most recent reports on the critical raw materials were presented concisely. A general evaluation was performed on the CCAs concept and the research directions. The advantages of using critical metals for particular applications were presented to acknowledge the difficulty in the substitution of such elements with other materials. In order to establish the level of involvement of CCAs in the reduction of critical metal in extreme environment applications, a presentation was made of the previous achievements in the field and the potential for the reduction of critical metal content through the use of multi-component compositions.

Powder Bed Fusion Additive Manufacturing Using Critical Raw Materials: A Review.

The term "critical raw materials" (CRMs) refers to various metals and nonmetals that are crucial to Europe's economic progress. Modern technologies enabling effective use and recyclability of CRMs are in critical demand for the EU industries. The use of CRMs, especially in the fields of biomedicine, aerospace, electric vehicles, and energy applications, is almost irreplaceable. Additive manufacturing (also referred to as 3D printing) is one of the key enabling technologies in the field of manufacturing which underpins the Fourth Industrial Revolution. 3D printing not only suppresses waste but also provides an efficient buy-to-fly ratio and possesses the potential to entirely change supply and distribution chains, significantly reducing costs and revolutionizing all logistics. This review provides comprehensive new insights into CRM-containing materials processed by modern additive manufacturing techniques and outlines the potential for increasing the efficiency of CRMs utilization and reducing the dependence on CRMs through wider industrial incorporation of AM and specifics of powder bed AM methods making them prime candidates for such developments.

Toxicological evaluation of MnAl based permanent magnets using different in vitro models.

Due to economic, environmental and geopolitical issues, the development of permanent magnets with a composition free of rare earth elements and with acceptable magnetic properties has been considered a priority by the international community, being MnAl based alloys amongst the most promising candidates. The aim of this work was to evaluate the toxicity of powders of two forms of newly developed MnAl(C) permanent magnets through exposure experiments applying three model organisms, using as a benchmark powders of a commercial rare-earth-containing magnet (Nd2Fe14B). For this purpose, the direct exposure to the different particles suspensions as well as to magnets leachates was evaluated. Both viability and oxidative stress assays were applied in an adenocarcinomic human alveolar basal epithelial cell line (A549) and in the yeast Saccharomyces cerevisiae, together with the bioluminescent inhibition assay in the Gram negative bacterium Vibrio fischeri. The obtained results indicate that MnAl(C) permanent magnets, in general terms, presented similar toxicity than the Nd magnet for the selected biological models under the studied conditions. Overall, the presented data provide, for the first time, an in vitro toxicity analysis of MnAl based magnets.

The rate of cochlear compression in a dolphin: a forward-masking evoked-potential study.

The "active" cochlear mechanism of hearing manifests in the cochlear compression. Investigations of compression in odontocetes help to determine the frequency limit of the active mechanism. The compression may be evaluated by comparison of low- and on-frequency masking. In a bottlenose dolphin, forward masking of auditory evoked potentials to tonal pips was investigated. Measurements were performed for test frequencies of 45 and 90 kHz. The low-frequency maskers were - 0.25 to - 0.75 oct relative the test. Masking efficiency was varied by masker-to-test delay variation from 2 to 20 ms, and masker levels at threshold (MLTs) were evaluated at each of the delays. It was assumed that low-frequency maskers were not subjected or little subjected to compression whereas on-frequency maskers were subjected equally to the test. Therefore, the compression rate was assessed as the slope of low-frequency MLT dependence on on-frequency MLT. For the 90-kHz test, the slopes were 0.63 and 0.18 dB/dB for masker of - 0.25 and - 0.5 oct, respectively. For the 45 kHz test, the slopes were 0.69 and 0.39 dB/dB for maskers of - 0.25 and - 0.5 oct. So, compression did not decay at the upper boundary of the hearing frequency range in the dolphin.

Large positive and negative Goos-Hänchen shifts near the surface plasmon resonance in subwavelength grating.

Diffraction of light of a visible spectral range by subwavelength metal gratings has been investigated experimentally and theoretically using rigorous electromagnetic calculations. It has been demonstrated that an effect of surface plasmon resonance (SPR), at which total absorption of light by metal grating can be observed, occurs under certain conditions. Large positive and negative Goos-Hänchen (GH) shifts occur near the SPR. It has been shown that the reflected beam is split into two parts, the relative powers of which depend on the incident beam width and the grating depth. The dependence of the GH shifts on the grating period and grating depth has been investigated for different incident beam widths. The high sensitivity of the GH shift on the incident angle of a light beam near the SPR has been demonstrated.

Spatial release from masking in a bottlenose dolphin Tursiops truncatus.

The impact of maskers on the receiving beam of a bottlenose dolphin, Tursiops truncatus, was investigated using the auditory evoked potential (AEP) method. The test signal was a train of tone pips with a 64 kHz carrier frequency. The stimulus produced AEPs as a sequence of waves replicating the pip rate: the rate following response (RFR). The masker was band-limited noise, with a passband of 45 to 90 kHz and a level of 105 dB re 1 µPa. Masker azimuths were at 0°, ±30°, ±60°, and ±90° relative to the head midline. The receiving beam was evaluated in terms of the RFR threshold dependence on the signal azimuth. The masked thresholds were higher than the baseline thresholds, which appeared mostly as a shift rather than a deformation in the receiving beam. The largest threshold shift appeared when the masker source was located in the most sensitive direction (zero azimuth); at lateral masker source positions, the threshold shift decreased. When the masker source was not at the head midline, the masked thresholds were higher at signal positions ipsilateral to the masker source than at positions contralateral to the masker source. The largest asymmetry was observed at the 30° masker azimuth in conjunction with the ±30° and ±120° signal azimuths; the asymmetries were 5.6 and 8.1 dB, respectively. This masking asymmetry was lower than expected from the previously found interaural intensity difference, which may be explained by the conflict between the test signal and the masker when it appeared at a binaural level of the auditory system.

Level-dependent masking of the auditory evoked responses in a dolphin: manifestation of the compressive nonlinearity.

At suprathreshold sound levels, interactions between masking noise and sound signals are liable to compressive nonlinearity in the auditory system. The compressive nonlinearity is a property of the "active" cochlear mechanism. It is not known whether this mechanism is capable to function at frequencies close to or above 100 kHz that are available to odontocetes (toothed whales, dolphins, and porpoises). This question may be answered by the use of the frequency-specific masking. Auditory evoked potentials to sound stimuli in a bottlenose dolphin, Tursiops truncatus, were recorded in the presence of simultaneous maskers. Stimulus frequencies were 45, 64, or 90 kHz. Maskers were on-frequency bandlimited noise or low-frequency noise of frequencies 0.25-1 oct below the stimulus frequency. The stimuli provoked responses as a series of brain-potential waves following the pip-train rate. For the on-frequency masker, the masker level at threshold dependence on the signal level was 1.1 dB/dB. For maskers of 1 oct below the stimulus, the dependence was 0.53-0.57 dB/dB. The data considered evidence for the compressive nonlinearity of responses to stimuli, and therefore, are indicative of the functioning of the active mechanism at frequencies up to 90 kHz.

Adaptation in the auditory system of a beluga whale: effect of adapting sound parameters.

The effects of adapting sounds (pip trains or pure tones) on auditory evoked potentials (the rate following response, RFR) were investigated in a beluga whale. During RFR acquisition, adapting signals lasting 128 ms each were alternated with test signals lasting 16 ms each; the test signal levels varied randomly. Adapting signals were trains of cosine-enveloped tone pips or pure tones. Pip rate varied with the envelope cosine cycle maintained at 0.125 of pip intervals and the cosine rise-fall time maintained at 0.0625 of pip intervals. Adapting signals shifted the amplitude-level function upward compared to the baseline (no adapting signal) function. The higher the adapting signal level was, the bigger the shift in the amplitude-level function was. The slower the pips were in the adapting signal, the smaller the adaptation effect was. A train of pips with a 0.0625-ms rise-fall time and 125 dB SPL shifted the function by 35-40 dB, whereas a train of pips with a 1-ms rise-fall time or a pure tone with the same SPL shifted the function by approximately 15 dB. The difference between the "fast" and "slow" adapting signals is supposed to be associated with their abilities to stimulate the auditory system in odontocetes.

Additive manufacturing to veterinary practice: recovery of bony defects after the osteosarcoma resection in canines.

The paper outlines the achievements and challenges in the additive manufacturing (AM) application to veterinary practice. The state-of-the-art in AM application to the veterinary surgery is presented, with the focus of AM for patient-specific implants manufacturing. It also provides critical discussion on some of the potential issues design and technology should overcome for wider and more effective implementation of additively manufactured parts in veterinary practices. Most of the discussions in present paper are related to the metallic implants, manufactured in this case using so-called powder bed additive manufacturing (PB-AM) in titanium alloy Ti-6AL-4V, and to the corresponding process of their design, manufacturing and implementation in veterinary surgery. Procedures of the implant design and individualization for veterinary surgery are illustrated basing on the four performed surgery cases with dog patients. Results of the replacement surgery in dogs indicate that individualized additively manufactured metallic implants significantly increase chances for successful recovery process, and AM techniques present a viable alternative to amputation in a large number of veterinary cases. The same time overcoming challenges of implant individualization in veterinary practice significantly contributes to the knowledge directly relevant to the modern medical practice. An experience from veterinary cases where organ-preserving surgery with 3D-printed patient-specific implants is performed provides a unique opportunity for future development of better human implants.

Selective electron beam melting of Al0.5CrMoNbTa0.5 high entropy alloys using elemental powder blend.

High Entropy Alloys (HEAs) is a novel promising class of multi-component materials which may demonstrate superior mechanical properties useful for high-temperature applications. Despite the high potential of HEAs, their production is complicated, using pre-alloyed powders in powder metallurgy route. This significantly complicates development and implementation of refractory BCC solid solution based HEAs. The present paper reports on experiments aiming at production of Al0.5CrMoNbTa0.5 multi-principle alloy using powder bed beam based additive manufacturing. Samples were manufactured using Selective Electron Beam Melting (SEBM) additive manufacturing technique from a blend of elemental powders aiming at achieving microstructure with high configurational entropy. Though it was not possible to achieve completely homogeneous microstructure, the as-printed material was composed of the zones with two multi-component solid solutions, which differed only by Al content confirming in situ alloying. The process parameters optimization was not carried out and the as-print material contained a notable amount of residual porosity. It was possible to reach lower porosity level using heat treatment at 1300 °C for 24 hours, however undesirable alloy composition changes took place. The main conclusion is that the production of the Al0.5CrMoNbTa0.5 multi-principle alloy from elemental powder blends using SEBM technique is achievable, but the process parameter optimization rather than post-process heat treatment should be performed to reduce the porosity of samples.