C\u3csub\u3e60\u3c/sub\u3e fullerenes selectively inhibit BK\u3csub\u3eCa\u3c/sub\u3e but not K\u3csub\u3ev\u3c/sub\u3e channels in pulmonary artery smooth muscle cells

© 2019 Elsevier Inc. Possessing unique physical and chemical properties, C60 fullerenes are arising as a potential nanotechnological tool that can strongly affect various biological processes. Recent molecular modeling studies have shown that C60 fullerenes can interact with ion channels, but there is lack of data about possible effects of C60 molecule on ion channels expressed in smooth muscle cells (SMC). Here we show both computationally and experimentally that water-soluble pristine C60 fullerene strongly inhibits the large conductance Ca2+-dependent K+ (BKCa), but not voltage-gated K+ (Kv) channels in pulmonary artery SMC. Both molecular docking simulations and analysis of single channel activity indicate that C60 fullerene blocks BKCa channel pore in its open state. In functional tests, C60 fullerene enhanced phenylephrine-induced contraction of pulmonary artery rings by about 25% and reduced endothelium-dependent acetylcholine-induced relaxation by up to 40%. These findings suggest a novel strategy for biomedical application of water-soluble pristine C60 fullerene in vascular dysfunction

Observation of Brownian Relaxation of Magnetic Nanoparticles using HTS SQUID

We analyzed the sensitivity of a separationless immunoassay scheme using functionalized magnetic nanoparticles (MNPs) and a sensitive HTS SQUID magnetometer. The signal of a 100 mu L sample at a concentration of 1 mg/mL and field of 7.5 nT was 20 m Phi(0). This makes it possible for the sensitivity to be within the range of 50 ng/mL at the required time of up to 100 s per a point in the frequency spectrum

Long-Read cDNA Sequencing Revealed Novel Expressed Genes and Dynamic Transcriptome Landscape of Triticale (x Triticosecale Wittmack) Seed at Different Developing Stages

Developing seed is a unique stage of plant development with highly dynamic changes in transcriptome. Here, we aimed to detect the novel previously unannotated, genes of the triticale (x Triticosecale Wittmack, AABBRR genome constitution) genome that are expressed during different stages and at different parts of the developing seed. For this, we carried out the Oxford Nanopore sequencing of cDNA obtained for middle (15 days post-anthesis, dpa) and late (20 dpa) stages of seed development. The obtained data together with our previous direct RNA sequencing of early stage (10 dpa) of seed development revealed 39,914 expressed genes including 7128 (17.6%) genes that were not previously annotated in A, B, and R genomes. The bioinformatic analysis showed that the identified genes belonged to long non-coding RNAs (lncRNAs), protein-coding RNAs, and TE-derived RNAs. The gene set analysis revealed the transcriptome dynamics during seed development with distinct patterns of over-represented gene functions in early and middle/late stages. We performed analysis of the lncRNA genes polymorphism and showed that the genes of some of the tested lncRNAs are indeed polymorphic in the triticale collection. Altogether, our results provide information on thousands of novel loci expressed during seed development that can be used as new targets for GWAS analysis, the marker-assisted breeding of triticale, and functional elucidation

Adiabatic superconducting cells for ultra-low-power artificial neural networks

We propose the concept of using superconducting quantum interferometers for the implementation of neural network algorithms with extremely low power dissipation. These adiabatic elements are Josephson cells with sigmoid- and Gaussian-like activation functions. We optimize their parameters for application in three-layer perceptron and radial basis function networks

Long-Read cDNA Sequencing Revealed Novel Expressed Genes and Dynamic Transcriptome Landscape of Triticale (x Triticosecale Wittmack) Seed at Different Developing Stages

Developing seed is a unique stage of plant development with highly dynamic changes in transcriptome. Here, we aimed to detect the novel previously unannotated, genes of the triticale (x Triticosecale Wittmack, AABBRR genome constitution) genome that are expressed during different stages and at different parts of the developing seed. For this, we carried out the Oxford Nanopore sequencing of cDNA obtained for middle (15 days post-anthesis, dpa) and late (20 dpa) stages of seed development. The obtained data together with our previous direct RNA sequencing of early stage (10 dpa) of seed development revealed 39,914 expressed genes including 7128 (17.6%) genes that were not previously annotated in A, B, and R genomes. The bioinformatic analysis showed that the identified genes belonged to long non-coding RNAs (lncRNAs), protein-coding RNAs, and TE-derived RNAs. The gene set analysis revealed the transcriptome dynamics during seed development with distinct patterns of over-represented gene functions in early and middle/late stages. We performed analysis of the lncRNA genes polymorphism and showed that the genes of some of the tested lncRNAs are indeed polymorphic in the triticale collection. Altogether, our results provide information on thousands of novel loci expressed during seed development that can be used as new targets for GWAS analysis, the marker-assisted breeding of triticale, and functional elucidation

Searching for a Needle in a Haystack: Cas9-Targeted Nanopore Sequencing and DNA Methylation Profiling of Full-Length Glutenin Genes in a Big Cereal Genome

Sequencing and epigenetic profiling of target genes in plants are important tasks with various applications ranging from marker design for plant breeding to the study of gene expression regulation. This is particularly interesting for plants with big genome size for which whole-genome sequencing can be time-consuming and costly. In this study, we asked whether recently proposed Cas9-targeted nanopore sequencing (nCATS) is efficient for target gene sequencing for plant species with big genome size. We applied nCATS to sequence the full-length glutenin genes (Glu-1Ax, Glu-1Bx and Glu-1By) and their promoters in hexaploid triticale (X Triticosecale, AABBRR, genome size is 24 Gb). We showed that while the target gene enrichment per se was quite high for the three glutenin genes (up to 645×), the sequencing depth that was achieved from two MinION flowcells was relatively low (5–17×). However, this sequencing depth was sufficient for various tasks including detection of InDels and single-nucleotide variations (SNPs), read phasing and methylation profiling. Using nCATS, we uncovered SNP and InDel variation of full-length glutenin genes providing useful information for marker design and deciphering of variation of individual Glu-1By alleles. Moreover, we demonstrated that glutenin genes possess a ‘gene-body’ methylation epigenetic profile with hypermethylated CDS part and hypomethylated promoter region. The obtained information raised an interesting question on the role of gene-body methylation in glutenin gene expression regulation. Taken together, our work disclosures the potential of the nCATS approach for sequencing of target genes in plants with big genome size

Performance and Mechanism of Photoelectrocatalytic Activity of MoSx/WO3 Heterostructures Obtained by Reactive Pulsed Laser Deposition for Water Splitting

This work studies the factors that affect the efficiency of the photoelectrochemical hydrogen evolution reaction (HER) using MoSx/WO3 nano-heterostructures obtained by reactive pulsed laser deposition (RPLD) on glass substrates covered with fluorinated tin oxide (FTO). Another focus of the research is the potential of MoSx nanofilms as a precursor for MoOz(S) nanofilms, which enhance the efficiency of the photo-activated oxygen evolution reaction (OER) using the MoOz(S)/WO3/FTO heterostructures. The nanocrystalline WO3 film was created by laser ablation of a W target in dry air at a substrate temperature of 420 °C. Amorphous MoSx nanofilms (2 ≤ x ≤ 12) were obtained by laser ablation of an Mo target in H2S gas of varied pressure at room temperature of the substrate. Studies of the energy band structures showed that for all MoSx/WO3/FTO samples, photo-activated HER in an acid solution proceeded through the Z-scheme. The highest photoelectrochemical HER efficiency (a photocurrent density ~1 mA/cm2 at a potential of ~0 V under Xe lamp illumination (~100 mW/cm2)) was found for porous MoS4.5 films containing the highest concentration of catalytically active sites attributed to S ligands. During the anodic posttreatment of porous MoSx nanofilms, MoOz(S) films with a narrow energy band gap were formed. The highest OER efficiency (a photocurrent density ~5.3 mA/cm2 at 1.6 V) was detected for MoOz(S)/WO3/FTO photoanodes that were prepared by posttreatment of the MoSx~3.2 precursor. The MoOz(S) film contributed to the effective photogeneration of electron–hole pairs that was followed by the transport of photoelectrons from MoOz(S) into the WO3 film and the effective participation of holes possessing strong oxidation ability in the OER on the surface of the MoOz(S) film

Genomic and Transcriptomic Survey Provides New Insight into the Organization and Transposition Activity of Highly Expressed LTR Retrotransposons of Sunflower (Helianthus annuus L.)

LTR retrotransposons (RTEs) play a crucial role in plant genome evolution and adaptation. Although RTEs are generally silenced in somatic plant tissues under non-stressed conditions, some expressed RTEs (exRTEs) escape genome defense mechanisms. As our understanding of exRTE organization in plants is rudimentary, we systematically surveyed the genomic and transcriptomic organization and mobilome (transposition) activity of sunflower (Helianthus annuus L.) exRTEs. We identified 44 transcribed RTEs in the sunflower genome and demonstrated their distinct genomic features: more recent insertion time, longer open reading frame (ORF) length, and smaller distance to neighboring genes. We showed that GAG-encoding ORFs are present at significantly higher frequencies in exRTEs, compared with non-expressed RTEs. Most exRTEs exhibit variation in copy number among sunflower cultivars and one exRTE Gagarin produces extrachromosomal circular DNA in seedling, demonstrating recent and ongoing transposition activity. Nanopore direct RNA sequencing of full-length RTE RNA revealed complex patterns of alternative splicing in RTE RNAs, resulting in isoforms that carry ORFs for distinct RTE proteins. Together, our study demonstrates that tens of expressed sunflower RTEs with specific genomic organization shape the hidden layer of the transcriptome, pointing to the evolution of specific strategies that circumvent existing genome defense mechanisms

The Gabbro–Granodiorite Magmatic Complex of the Kronotsky Paleoarc (Eastern Kamchatka): Composition, Age, and Tectonic Position

New U‒Pb (LA-ICP-MS) geochronological data have been obtained on accessory zircons from granodiorites and on detrital zircons from stream-sediment samples from the Shipunsky massif in the Eastern Kamchatka region. The age of accessory zircons from amphibole–biotite granodiorites has been estimated at 49–44 Ma. Detrital zircons have the Late Paleocene–Early Eocene age from ~57 to ~49 Ma. Based on the geological and geochronological data, the massif was formed in two stages: a gabbroid intrusion (56‒51 Ma) and the quartz diorite-granodiorite intrusion (49‒44 Ma). In terms of the petrographic and geochemical characteristics of the Upper Cretaceous–Eocene volcanic rocks in the Shipunsky Peninsula and granitoids in the Shipunsky massif, they were formed in the suprasubduction setting. The Shipunsky granitoids belong to the I-type granites. The Shipunsky massif was formed as a part of the Kronotsky intraoceanic paleoarc during the Paleocene–Eocene in two stages. The southern segment of the Kronotsky paleoarc collided with the Kamchatka continental margin and the deformed rocks of this massif were brought to the surface