Tubular cell and keratinocyte single-cell transcriptomics applied to lupus nephritis reveal type I IFN and fibrosis relevant pathways.

The molecular and cellular processes that lead to renal damage and to the heterogeneity of lupus nephritis (LN) are not well understood. We applied single-cell RNA sequencing (scRNA-seq) to renal biopsies from patients with LN and evaluated skin biopsies as a potential source of diagnostic and prognostic markers of renal disease. Type I interferon (IFN)-response signatures in tubular cells and keratinocytes distinguished patients with LN from healthy control subjects. Moreover, a high IFN-response signature and fibrotic signature in tubular cells were each associated with failure to respond to treatment. Analysis of tubular cells from patients with proliferative, membranous and mixed LN indicated pathways relevant to inflammation and fibrosis, which offer insight into their histologic differences. In summary, we applied scRNA-seq to LN to deconstruct its heterogeneity and identify novel targets for personalized approaches to therapy

Safety of procuring research tissue during a clinically indicated kidney biopsy from patients with lupus: data from the Accelerating Medicines Partnership RA/SLE Network

Objectives In lupus nephritis the pathological diagnosis from tissue retrieved during kidney biopsy drives treatment and management. Despite recent approval of new drugs, complete remission rates remain well under aspirational levels, necessitating identification of new therapeutic targets by greater dissection of the pathways to tissue inflammation and injury. This study assessed the safety of kidney biopsies in patients with SLE enrolled in the Accelerating Medicines Partnership, a consortium formed to molecularly deconstruct nephritis.Methods 475 patients with SLE across 15 clinical sites in the USA consented to obtain tissue for research purposes during a clinically indicated kidney biopsy. Adverse events (AEs) were documented for 30 days following the procedure and were determined to be related or unrelated by all site investigators. Serious AEs were defined according to the National Institutes of Health reporting guidelines.Results 34 patients (7.2%) experienced a procedure-related AE: 30 with haematoma, 2 with jets, 1 with pain and 1 with an arteriovenous fistula. Eighteen (3.8%) experienced a serious AE requiring hospitalisation; four patients (0.8%) required a blood transfusion related to the kidney biopsy. At one site where the number of cores retrieved during the biopsy was recorded, the mean was 3.4 for those who experienced a related AE (n=9) and 3.07 for those who did not experience any AE (n=140). All related AEs resolved.Conclusions Procurement of research tissue should be considered feasible, accompanied by a complication risk likely no greater than that incurred for standard clinical purposes. In the quest for targeted treatments personalised based on molecular findings, enhanced diagnostics beyond histology will likely be required

Modulation of microRNA function by synthetic ribozymes

MicroRNAs (miRNAs) are now recognized as one of the major class of gene regulatory molecules in eukaryotic cells. Aberrant miRNA expression has been implicated in many human diseases. Herein, we exploit the site-specific cleavage ability of hammerhead ribozymes to design synthetic ribozymes and establish that they can cleave miRNA, thereby inhibiting miRNA function. We also used modified ribozymes where a 3′-3′-linked nucleotide “cap” (inverted T) was added and few ribonucleotides were changed to 2′-O-methyl nucleotides. The modified ribozyme was more efficient at inhibiting miR-21 than the wild type ribozyme

Synthesis and Biological Evaluation of Some Substituted Benzimidazole Derivatives

In the current research work, the title compounds 5-ethoxy-benzimidazole, were synthesized by nitration of phenacetin with concentrated nitric acid it gives N-(2-nitro-5-ethoxyphenyl) acetamide (I), which on reduction with alcohol gives 5-ethoxy-2-nitroaniline (II). Reaction of hydrazine hydrate with 5-ethoxy-2-nitroaniline produced 5-ethoxy ortho phenylene diamine (III). The substituted acids reacted with 5-ethoxy ortho phenylene diamine then yielded the corresponding 5-ethoxy-benzimidazole (IV). The identification and characterization of the synthesized compounds were carried out by Elemental analysis, melting point, Thin Layer Chromatography, FT-IR, NMR and Mass data. The synthesized compounds were evaluated for anti-tubercular activity. The test compounds were subjected to in vitro screening by the tube dilution technique employing the human virulent H37RV strain of M. tuberculosis. The test compounds IVa, IVc and IVd showed significant anti-tubercular activity against H37RV strain of Mycobacterium tuberculosis. The minimum inhibitory concentration (MIC) values were found in the range of 0.8 to 12.5 μg/mL compared with the standard drugs Isoniazid

Mammalian miRNA curation through next-generation sequencing

Characteristic small RNA biogenesis processing patterns are used for the discovery of novel microRNAs (miRNAs) from next-generation sequencing data. Here, we highlight and discuss key criteria for mammalian – specifically human – miRNA database curation based on small RNA sequencing data. Sequence reads obtained from small RNA cDNA libraries are aligned to reference genomic regions, and miRNA genes are revealed by their distinct read length and bimodal read frequency distribution, the predicted secondary structure of the deduced miRNA stem-loop precursor molecule, and, to a lesser degree, based on evolutionary conservation of small RNAs from other vertebrates. Properly curated miRNA databases are an important resource for investigators interested in miRNA biology, diagnostics, and therapeutics

Specificity of RSG-1.2 Peptide Binding to RRE-IIB RNA Element of HIV-1 over Rev Peptide Is Mainly Enthalpic in Origin

Rev is an essential HIV-1 regulatory protein which binds to the Rev responsive element (RRE) present within the env gene of HIV-1 RNA genome. This binding facilitates the transport of the RNA to the cytoplasm, which in turn triggers the switch between viral latency and active viral replication. Essential components of this complex have been localized to a minimal arginine rich Rev peptide and stem IIB region of RRE. A synthetic peptide known as RSG-1.2 binds with high binding affinity and specificity to the RRE-IIB than the Rev peptide, however the thermodynamic basis of this specificity has not yet been addressed. The present study aims to probe the thermodynamic origin of this specificity of RSG-1.2 over Rev Peptide for RRE-IIB. The temperature dependent melting studies show that RSG-1.2 binding stabilizes the RRE structure significantly (DT m = 4.3uC), in contrast to Rev binding. Interestingly the thermodynamic signatures of the binding have also been found to be different for both the peptides. At pH 7.5, RSG-1.2 binds RRE-IIB with a Ka = 16.260.6610 7 M 21 where enthalpic change DH = 213.960.1 kcal/mol is the main driving force with limited unfavorable contribution from entropic change TDS = 22.860.1 kcal/mol. A large part of DH may be due to specific stacking between U72 and Arg15. In contrast binding of Rev (K a = 3.160.4610 7 M 21) is driven mainly by entropy (DH = 0 kcal/mol and TDS = 10.260.2 kcal/mol) which arises fro

Conjugated Polymeric Liposomes: A Hybrid Carrier for Contemporary Drug Delivery

Liposomes are artificial vesicles encapsulating the drug moiety. The structural adaptability of liposomes has been employed to make them drug carriers for smart delivery systems, improving bioavailability, stability, target delivery, etc. However, conventional liposomes have some drawbacks, such as limited payload, shorter in vivo circulatory lifespan, unregulated releasing properties, rapid clearance from bloodstream, etc. Polymeric modification of the liposomes addressed and effectively overcame all the drawbacks of conventional liposomes. Polymeric materials offer indefinite structural diversity, thus a substantial portion of the materials has been employed for drug-targeting methods and controlled drug release. Conjugation of liposomes and polymers develops a hybrid vesicle with intermediary physicochemical and stimulus responsive properties (pH, temperature, etc.). The reliability of liposomes with respect to pH, nature of drug moiety, enzyme, and immune response can be strengthened by polymers. Polymer modified liposomes also enhance the pharmacokinetic and pharmacodynamic profile of the drug moiety. The form of polymer, cross-linking agent, interaction, and bonding used during polymerized modification of liposomes all have an impact on their activity. According to the extensive review of the literature that is accessible in the different data sources, research in this field is proactively involved in the synthesis of newer polymeric materials, and the supramolecular structuring of the different chemicals

Conjugated Polymeric Liposomes: A Hybrid Carrier for Contemporary Drug Delivery

Liposomes are artificial vesicles encapsulating the drug moiety. The structural adaptability of liposomes has been employed to make them drug carriers for smart delivery systems, improving bioavailability, stability, target delivery, etc. However, conventional liposomes have some drawbacks, such as limited payload, shorter in vivo circulatory lifespan, unregulated releasing properties, rapid clearance from bloodstream, etc. Polymeric modification of the liposomes addressed and effectively overcame all the drawbacks of conventional liposomes. Polymeric materials offer indefinite structural diversity, thus a substantial portion of the materials has been employed for drug-targeting methods and controlled drug release. Conjugation of liposomes and polymers develops a hybrid vesicle with intermediary physicochemical and stimulus responsive properties (pH, temperature, etc.). The reliability of liposomes with respect to pH, nature of drug moiety, enzyme, and immune response can be strengthened by polymers. Polymer modified liposomes also enhance the pharmacokinetic and pharmacodynamic profile of the drug moiety. The form of polymer, cross-linking agent, interaction, and bonding used during polymerized modification of liposomes all have an impact on their activity. According to the extensive review of the literature that is accessible in the different data sources, research in this field is proactively involved in the synthesis of newer polymeric materials, and the supramolecular structuring of the different chemicals

Screening of Rubiaceae and Apocynaceae extracts for mosquito larvicidal potential

<div><p>Rubiaceae and Apocynaceae families are well known for the expression of cyclotides having insecticidal properties. Leaves and flowers extracts of plants from the families Rubiaceae (<i>Ixora coccinea</i>) and Apocynaceae (<i>Allamanda violacea</i>) were evaluated for mosquito larvicidal effect against early IVth instars of <i>Aedes aegypti</i> and <i>Anopheles stephensi</i>. Two forms of plant extracts, one untreated and the other treated with heat and proteolytic enzyme were used for assay. After primary assay, the extract showing more than 50% inhibition was further used for quantification purpose. LC₅₀ and LC₉₀ values of all the extracts were found to be reduced with the treated form. Phytochemical analysis of plant extracts was performed. Primary confirmation for the presence of cyclotides was done by Lowry test, thin layer chromatography and haemolytic assay. This novel approach merits use of plant extracts in mosquito control programmes.</p></div

Antagonism of microRNA function in zebrafish embryos by using Locked Nucleic Acid enzymes (LNAzymes)

MicroRNAs (miRNAs) have crucial functions in many cellular processes, such as differentiation, proliferation and apoptosis; aberrant expression of miRNAs has been linked to human diseases, including cancer. Tools that allow specific and efficient knockdown of miRNAs would be of immense importance for exploring miRNA function. Zebrafish serves as an excellent vertebrate model system to understand the functions of miRNAs involved in a variety of biological processes. We designed and employed a strategy based on locked nucleic acid enzymes (LNAzymes) for in vivo knockdown of miRNA in zebrafish embryos. We demonstrate that LNAzyme can efficiently knockdown miRNAs with minimal toxicity to the zebrafish embryos