LncRNA VEAL2 regulates PRKCB2 to modulate endothelial permeability in diabetic retinopathy

Long non‐coding RNAs (lncRNAs) are emerging as key regulators of endothelial cell function. Here, we investigated the role of a novel vascular endothelial‐associated lncRNA (VEAL2) in regulating endothelial permeability. Precise editing of veal2 loci in zebrafish (veal2 ^gib005Δ8/+) induced cranial hemorrhage. In vitro and in vivo studies revealed that veal2 competes with diacylglycerol for interaction with protein kinase C beta‐b (Prkcbb) and regulates its kinase activity. Using PRKCB2 as bait, we identified functional ortholog of veal2 in humans from HUVECs and named it as VEAL2. Overexpression and knockdown of VEAL2 affected tubulogenesis and permeability in HUVECs. VEAL2 was differentially expressed in choroid tissue in eye and blood from patients with diabetic retinopathy, a disease where PRKCB2 is known to be hyperactivated. Further, VEAL2 could rescue the effects of PRKCB2‐mediated turnover of endothelial junctional proteins thus reducing hyperpermeability in hyperglycemic HUVEC model of diabetic retinopathy. Based on evidence from zebrafish and hyperglycemic HUVEC models and diabetic retinopathy patients, we report a hitherto unknown VEAL2 lncRNA‐mediated regulation of PRKCB2, for modulating junctional dynamics and maintenance of endothelial permeability.     

Hybrid Polymer Electrolyte Encased Cathode Particles Interface-Based Core–Shell Structure for High-Performance Room Temperature All-Solid-State Batteries

A smooth interfacial contact between electrode and electrolyte, alleviation of dendrite formation, low internal resistance, and preparation of thin electrolyte (<20 µm) are the key challenging tasks in the practical application of Li₇La₃Zr₂O₁₂ (LLZO)-based solid-state batteries (SSBs). This paper develops a unique strategy to reduce interfacial resistance by designing an interface-based core–shell structure via direct integration of Al-LLZO ceramic nanofibers incorporated poly(vinylidene fluoride)/LiTFSI on the surface of a porous cathode electrode (HPEIC). This yields an ultrathin solid polymer electrolyte with a thickness of 7 µm. The integrated HPEIC/Li SSB with LiFePO₄/C exhibits an initial specific capacity of 166 mAh g⁻¹ at 0.1 C and 159 mAh g⁻¹ with capacity retention of 100% after 120 cycles at 0.5 C (25 °C). The HPEIC/Li SSB with LiNi_0.8Mn_0.1Co_0.1O₂ cathode delivers a good discharge capacity of 134 mAh g⁻¹ after 120 cycles at 0.5 C. The rational design of interface-based core–shell structure outperforms the conventional assembly of solid-state cells using free-standing solid electrolytes in specific capacity, internal resistance, and rate performance. The proposed strategy is simple, cost-effective, robust, and scalable manufacturing, which is essential for the practical applicability of SSBs.     

Comparative Whole-Genome Analysis of Clinical Isolates Reveals Characteristic Architecture of Mycobacterium tuberculosis Pangenome

The tubercle complex consists of closely related mycobacterium species which appear to be variants of a single species. Comparative genome analysis of different strains could provide useful clues and insights into the genetic diversity of the species. We integrated genome assemblies of 96 strains from Mycobacterium tuberculosis complex (MTBC), which included 8 Indian clinical isolates sequenced and assembled in this study, to understand its pangenome architecture. We predicted genes for all the 96 strains and clustered their respective CDSs into homologous gene clusters (HGCs) to reveal a hard-core, soft-core and accessory genome component of MTBC. The hard-core (HGCs shared amongst 100% of the strains) was comprised of 2,066 gene clusters whereas the soft-core (HGCs shared amongst at least 95% of the strains) comprised of 3,374 gene clusters. The change in the core and accessory genome components when observed as a function of their size revealed that MTBC has an open pangenome. We identified 74 HGCs that were absent from reference strains H37Rv and H37Ra but were present in most of clinical isolates. We report PCR validation on 9 candidate genes depicting 7 genes completely absent from H37Rv and H37Ra whereas 2 genes shared partial homology with them accounting to probable insertion and deletion events. The pangenome approach is a promising tool for studying strain specific genetic differences occurring within species. We also suggest that since selecting appropriate target genes for typing purposes requires the expected target gene be present in all isolates being typed, therefore estimating the core-component of the species becomes a subject of prime importance.     

Impact of Fullerene on the Photophysics of Ternary Small Molecule Organic Solar Cells

Ternary organic solar cells (OSCs) are among the best‐performing organic photovoltaic devices to date, largely due to the recent development of nonfullerene acceptors. However, fullerene molecules still play an important role in ternary OSC systems, since, for reasons not well understood, they often improve the device performance, despite their lack of absorption. Here, the photophysics of a prototypical ternary small‐molecule OSC blend composed of the donor DR3, the nonfullerene acceptor ICC6, and the fullerene derivative PC₇₁BM is studied by ultrafast spectroscopy. Surprisingly, it is found that after excitation of PC₇₁BM, ultrafast singlet energy transfer to ICC6 competes efficiently with charge transfer. Subsequently, singlets on ICC6 undergo hole transfer to DR3, resulting in free charge generation. Interestingly, PC₇₁BM improves indirectly the electron mobility of the ternary blend, while electrons reside predominantly in ICC6 domains as indicated by fast spectroscopy. The improved mobility facilitates charge carrier extraction, in turn leading to higher device efficiencies of the ternary compared to binary solar cells. Using the (photo)physical parameters obtained from (transient) spectroscopy and charge transport measurements, the device's current–voltage characteristics are simulated and it is demonstrated that the parameters accurately reproduce the experimentally measured device performance.     

Screening Currency Notes for Microbial Pathogens and Antibiotic Resistance Genes Using a Shotgun Metagenomic Approach

Fomites are a well-known source of microbial infections and previous studies have provided insights into the sojourning microbiome of fomites from various sources. Paper currency notes are one of the most commonly exchanged objects and its potential to transmit pathogenic organisms has been well recognized. Approaches to identify the microbiome associated with paper currency notes have been largely limited to culture dependent approaches. Subsequent studies portrayed the use of 16S ribosomal RNA based approaches which provided insights into the taxonomical distribution of the microbiome. However, recent techniques including shotgun sequencing provides resolution at gene level and enable estimation of their copy numbers in the metagenome. We investigated the microbiome of Indian paper currency notes using a shotgun metagenome sequencing approach. Metagenomic DNA isolated from samples of frequently circulated denominations of Indian currency notes were sequenced using Illumina Hiseq sequencer. Analysis of the data revealed presence of species belonging to both eukaryotic and prokaryotic genera. The taxonomic distribution at kingdom level revealed contigs mapping to eukaryota (70%), bacteria (9%), viruses and archae (~1%). We identified 78 pathogens including Staphylococcus aureus, Corynebacterium glutamicum, Enterococcus faecalis, and 75 cellulose degrading organisms including Acidothermus cellulolyticus, Cellulomonas flavigena and Ruminococcus albus. Additionally, 78 antibiotic resistance genes were identified and 18 of these were found in all the samples. Furthermore, six out of 78 pathogens harbored at least one of the 18 common antibiotic resistance genes. To the best of our knowledge, this is the first report of shotgun metagenome sequence dataset of paper currency notes, which can be useful for future applications including as bio-surveillance of exchangeable fomites for infectious agents.     

Loss of function mutation in the P2X7, a ligand-gated ion channel gene associated with hypertrophic cardiomyopathy

Purinergic Signalling - Hypertrophic cardiomyopathy (HCM) is an inherited heart failure condition, mostly found to have genetic abnormalities, and is a leading cause of sudden death in young...     

Aptamer-Assisted Detection of the Altered Expression of Estrogen Receptor Alpha in Human Breast Cancer

An increase in the expression of estrogen receptors (ER) and the expanded population of ER-positive cells are two common phenotypes of breast cancer. Detection of the aberrantly expressed ERα in breast cancer is carried out using ERα-antibodies and radiolabelled ligands to make decisions about cancer treatment and targeted therapy. Capitalizing on the beneficial advantages of aptamer over the conventional antibody or radiolabelled ligand, we have identified a DNA aptamer that selectively binds and facilitates the detection of ERα in human breast cancer tissue sections. The aptamer is identified using the high throughput sequencing assisted SELEX screening. Biophysical characterization confirms the binding and formation of a thermodynamically stable complex between the identified DNA aptamer (ERaptD4) and ERα (Ka = 1.55±0.298×10⁸ M^-1; ΔH = 4.32×10⁴±801.1 cal/mol; ΔS = -108 cal/mol/deg). Interestingly, the specificity measurements suggest that the ERaptD4 internalizes into ERα-positive breast cancer cells in a target-selective manner and localizes specifically in the nuclear region. To harness these characteristics of ERaptD4 for detection of ERα expression in breast cancer samples, we performed the aptamer-assisted histochemical analysis of ERα in tissue samples from breast cancer patients. The results were validated by performing the immunohistochemistry on same samples with an ERα-antibody. We found that the two methods agree strongly in assay output (kappa value = 0.930, p-value <0.05 for strong ERα positive and the ERα negative samples; kappa value = 0.823, p-value <0.05 for the weak/moderate ER+ve samples, n = 20). Further, the aptamer stain the ERα-positive cells in breast tissues without cross-reacting to ERα-deficient fibroblasts, adipocytes, or the inflammatory cells. Our results demonstrate a significant consistency in the aptamer-assisted detection of ERα in strong ERα positive, moderate ERα positive and ERα negative breast cancer tissues. We anticipate that the ERaptD4 aptamer targeting ERα may potentially be used for an efficient grading of ERα expression in cancer tissues.     

Charge and Triplet Exciton Generation in Neat PC70BM Films and Hybrid CuSCN:PC70BM Solar Cells

Organic solar cells that use only fullerenes as the photoactive material exhibit poor exciton‐to‐charge conversion efficiencies, resulting in low internal quantum efficiencies (IQE). However, the IQE can be greatly improved, when copper(I) thiocyanate (CuSCN) is used as a carrier‐selective interlayer between the phenyl‐C70‐butyric acid methyl ester (PC₇₀BM) layer and the anode. Efficiencies of ≈5.4% have recently been reported for optimized CuSCN:PC₇₀BM (1:3)‐mesostructured heterojunctions, yet the reasons causing the efficiency boost remain unclear. Here, transient absorption (TA) spectroscopy is used to demonstrate that CuSCN does not only act as a carrier‐selective electrode layer, but also facilitates fullerene exciton dissociation and hole transfer at the interface with PC₇₀BM. While intrinsic charge generation in neat PC₇₀BM films proceeds with low yield, hybrid films exhibit much improved exciton dissociation due to the presence of abundant interfaces. Triplet generation with a rate proportional to the product of singlet and charge concentrations is observed in neat PC₇₀BM films, implying a charge–singlet spin exchange mechanism, while in hybrid films, this mechanism is absent and triplet formation is a consequence of nongeminate recombination of free charges. At low carrier concentrations, the fraction of charges outweighs the population of triplets, leading to respectable device efficiencies under one sun illumination.