cNTnC and fYTnC2, Genetically Encoded Green Calcium Indicators Based on Troponin C from Fast Animals

NTnC-like green fluorescent genetically encoded calcium indicators (GECIs) with two calcium ion binding sites were constructed using the insertion of truncated troponin C (TnC) from Opsanus tau into green fluorescent proteins (GFPs). These GECIs are small proteins containing the N- and C-termini of GFP; they exert a limited effect on the cellular free calcium ion concentration; and in contrast to calmodulin-based calcium indicators they lack undesired interactions with intracellular proteins in neurons. The available TnC-based NTnC or YTnC GECIs had either an inverted response and high brightness but a limited dynamic range or a positive response and fast kinetics in neurons but lower brightness and an enhanced but still limited dF/F dynamic range. Here, we solved the crystal structure of NTnC at 2.5 Å resolution. Based on this structure, we developed positive NTnC2 and inverted iNTnC2 GECIs with a large dF/F dynamic range in vitro but very slow rise and decay kinetics in neurons. To overcome their slow responsiveness, we swapped TnC from O. tau in NTnC2 with truncated troponin C proteins from the muscles of fast animals, namely, the falcon, hummingbird, cheetah, bat, rattlesnake, and ant, and then optimized the resulting constructs using directed molecular evolution. Characterization of the engineered variants using purified proteins, mammalian cells, and neuronal cultures revealed cNTnC GECI with truncated TnC from Calypte anna (hummingbird) to have the largest dF/F fluorescence response and fast dissociation kinetics in neuronal cultures. In addition, based on the insertion of truncated TnCs from fast animals into YTnC2, we developed fYTnC2 GECI with TnC from Falco peregrinus (falcon). The purified proteins cNTnC and fYTnC2 had 8- and 6-fold higher molecular brightness and 7- and 6-fold larger dF/F responses to the increase in Ca²⁺ ion concentration than YTnC, respectively. cNTnC GECI was also 4-fold more photostable than YTnC and fYTnC2 GECIs. Finally, we assessed the developed GECIs in primary mouse neuronal cultures stimulated with an external electric field; in these conditions, cNTnC had a 2.4-fold higher dF/F fluorescence response than YTnC and fYTnC2 and was the same or slightly slower (1.4-fold) than fYTnC2 and YTnC in the rise and decay half-times, respectively.     

The Impact of Backbone Fluorination and Side-Chain Position in Thiophene-Benzothiadiazole-Based Hole-Transport Materials on the Performance and Stability of Perovskite Solar Cells

Perovskite solar cells (PSCs) currently reach high efficiencies, while their insufficient stability remains an obstacle to their technological commercialization. The introduction of hole-transport materials (HTMs) into the device structure is a key approach for enhancing the efficiency and stability of devices. However, currently, the influence of the HTM structure or properties on the characteristics and operational stability of PSCs remains insufficiently studied. Herein, we present four novel push-pull small molecules, H1-4, with alternating thiophene and benzothiadiazole or fluorine-loaded benzothiadiazole units, which contain branched and linear alkyl chains in the different positions of terminal thiophenes to evaluate the impact of HTM structure on PSC performance. It is demonstrated that minor changes in the structure of HTMs significantly influence their behavior in thin films. In particular, H3 organizes into highly ordered lamellar structures in thin films, which proves to be crucial in boosting the efficiency and stability of PSCs. The presented results shed light on the crucial role of the HTM structure and the morphology of films in the performance of PSCs.     

Combined In Silico,Ex Vivo,and In Vivo Assessment of L-17, a Thiadiazine Derivative with Putative Neuro- and Cardioprotective and Antidepressant Effects

Depression associated with poor general medical condition, such as post-stroke (PSD) or post-myocardial infarction (PMID) depression, is characterized by resistance to classical antidepressants. Special treatment strategies should thus be developed for these conditions. Our study aims to investigate the mechanism of action of 2-morpholino-5-phenyl-6H-1,3,4-thiadiazine, hydrobromide (L-17), a recently designed thiadiazine derivative with putative neuro- and cardioprotective and antidepressant-like effects, using combined in silico (for prediction of the molecular binding mechanisms), ex vivo (for assessment of the neural excitability using c-Fos immunocytochemistry), and in vivo (for direct examination of the neuronal excitability) methodological approaches. We found that the predicted binding affinities of L-17 to serotonin (5-HT) transporter (SERT) and 5-HT₃ and 5-HT_1A receptors are compatible with selective 5-HT serotonin reuptake inhibitors (SSRIs) and antagonists of 5-HT₃ and 5-HT_1A receptors, respectively. L-17 robustly increased c-Fos immunoreactivity in the amygdala and decreased it in the hippocampus. L-17 dose-dependently inhibited 5-HT neurons of the dorsal raphe nucleus; this inhibition was partially reversed by the 5-HT_1A antagonist WAY100135. We suggest that L-17 is a potent 5-HT reuptake inhibitor and partial antagonist of 5-HT₃ and 5-HT_1A receptors; the effects of L-17 on amygdaloid and hippocampal excitability might be mediated via 5-HT, and putatively mediate the antidepressant-like effects of this drug. Since L-17 also possesses neuro- and cardioprotective properties, it can be beneficial in PSD and PMID. Combined in silico predictions with ex vivo neurochemical and in vivo electrophysiological assessments might be a useful strategy for early assessment of the efficacy and neural mechanism of action of novel CNS drugs.     

Nanopore Sequencing for De Novo Bacterial Genome Assembly and Search for Single-Nucleotide Polymorphism

Nanopore sequencing (ONT) is a new and rapidly developing method for determining nucleotide sequences in DNA and RNA. It serves the ability to obtain long reads of thousands of nucleotides without assembly and amplification during sequencing compared to next-generation sequencing. Nanopore sequencing can help for determination of genetic changes leading to antibiotics resistance. This study presents the application of ONT technology in the assembly of an E. coli genome characterized by a deletion of the tolC gene and known single-nucleotide variations leading to antibiotic resistance, in the absence of a reference genome. We performed benchmark studies to determine minimum coverage depth to obtain a complete genome, depending on the quality of the ONT data. A comparison of existing programs was carried out. It was shown that the Flye program demonstrates plausible assembly results relative to others (Shasta, Canu, and Necat). The required coverage depth for successful assembly strongly depends on the size of reads. When using high-quality samples with an average read length of 8 Kbp or more, the coverage depth of 30× is sufficient to assemble the complete genome de novo and reliably determine single-nucleotide variations in it. For samples with shorter reads with mean lengths of 2 Kbp, a higher coverage depth of 50× is required. Avoiding of mechanical mixing is obligatory for samples preparation. Nanopore sequencing can be used alone to determine antibiotics-resistant genetic features of bacterial strains.     

Synthesis of -leucine nanoparticles via physical vapor deposition at varying saturation conditions

Preparation of L-leucine nanoparticles by a process based on physical vapor deposition has been presented. In an aerosol flow reactor method, aqueous L-leucine droplets were first dried followed by the sublimation of L-leucine to produce vapor that upon vapor deposition resulted in L-leucine nanoparticles with size ranging from 40 to 200 nm. Onset temperature for the sublimation of L-leucine at concentrations from 0.02 to increased from 135 to , respectively. The formation of nanoparticles was initiated in three different ways: (i) via droplet drying, (ii) via heterogeneous nucleation of L-leucine vapor on solid L-leucine particles, and (iii) via homogeneous nucleation of L-leucine vapor to form new-born nanoparticles. Consequently, the saturation conditions of L-leucine vapor in the reactor determined the resulting particle size, size distribution and number concentration, those depending very much on nucleation mode. In general, the both nucleation modes produced narrow size distributions, that is, geometric standard deviation (GSD) was <1.8 although the number concentration increased with the increased amount of L-leucine vapor. Upon desublimation and vapor deposition, L-leucine formed leafy crystals whose size was the largest when produced from the heated section at the vicinity of the onset temperature and the smallest far above the onset temperature. All the particles prepared in the conditions (i)–(iii) were crystalline. However, X-ray diffraction analysis showed preferential direction for crystal growth according to the way of particle formation.     

A Review of Acetylene Compounds as Inhibitors of Acid Corrosion of Iron

Literature data on acetylene compounds as inhibitors of acid corrosion of iron were reviewed. Inhibition mechanism–structure correlations were proposed for classification of acetylene compounds.     

A new inhibitor of steel corrosion in sufuric acid

A new effective inhibitor of steel corrosion in sulfuric acid, QAS-K1, from the class of quaternary ammonium salts was developed. The effects of the new inhibitor on corrosion of St3 steel in 2 M H₂SO₄ were studied by gravimetry at 25 and 60°C. In the individual state, QAS-K1 proved to be more effective than the industrial inhibitor catamine AB from a similar class. Anionic (5.0 mmol/l KI, 50.0 mmol/l KBr, and 5.0 mmol/l KCNS) and molecular (0.44 mmol/l diphenylthiourea (DPTU) and 0.60 mmol/l captax) additions increased the efficiency of the new inhibitor. QAS-K1 in mixtures with KI, KBr, DPTU, and captax appeared to be more effective in hindering steel corrosion in H₂SO₄ than similar mixtures based on catamine AB and tribenzylethanolammonium chloride. Mixtures based on QAS-K1 show a predominantly antagonistic action of the components. Original Russian Text ? Ya.G. Avdeev, P.A. Belinskii, Yu.I. Kuznetsov, O.O. Zel’, 2007, published in Zashchita Metallov, 2007, Vol. 43, No. 6, pp. 626–629.     

X‐ray micro‐computed tomography analysis of accumulated corrosion products in deep‐water shipwrecks

Accumulated corrosion products from two different shipwrecks which had lain on the seabed (2.5 km depth) for 73 years were systematically analysed by three‐dimensional imaging at high resolution using X‐ray micro‐computed tomography. Complementary surface and chemical characterization experiments were conducted to identify the morphological structure of the corrosion products. Goethite was observed as the main corrosion phase found in both the wreck's corrosion products. However, other corrosion products such as silica, lepidocrocite, maghemite, magnetite, benyacarite, jarosite and amorphous materials were noticed using Fourier transform infrared spectroscopy, Raman spectroscopy and X‐ray diffraction. In addition, mineralized microbial structures were also observed as significant constituents of the corrosion products. However, there were significant differences between samples from the two shipwrecks including porosity, distribution and volume percent of the corrosion products components. The mechanism of different corrosion products formation was proposed and discussed in detail.     

Piezoelectric K0.5Na0.5NbO3 Ceramics Textured Using Needlelike K0.5Na0.5NbO3 Templates

Improved performance by texturing has become attractive in the field of lead‐free ferroelectrics, but the effect depends heavily on the degree of texture, type of preferred orientation, and whether the material is a rotator or extender ferroelectric. Here, we report on successful texturing of K_0.5Na_0.5NbO₃ (KNN) ceramics by alignment of needlelike KNN templates in a matrix of KNN powder using tape casting. Homotemplated grain growth of the needles was confirmed during sintering, resulting in a high degree of texture parallel to the tape casting direction (TCD) and the aligned needles. The texture significantly improved the piezoelectric response parallel to the tape cast direction, corresponding to the direction of the strongest <001>_pc orientation, while the response normal to the tape cast plane was lower than for a nontextured KNN. In situ X‐ray diffraction during electric field application revealed that non‐180° domain reorientation was enhanced by an order of magnitude in the TCD, compared to the direction normal to the tape cast plane and in the nontextured ceramic. The effect of texture in KNN is discussed with respect to possible rotator ferroelectric properties of KNN.