The effects of various parameters of synthesis on the structure formation of layered double hydroxides (LDH) of the general formula M3Al(OH)8[(CO3)1/2 · mH2O], where M = Zn2+, Mg2+, or Cu2+, were...

Anti-cancer therapy based on oncolytic viruses (OVs) is a targeted approach that takes advantage of OVs’ ability to selectively infect and replicate in tumor cells, activate the host immune response, and destroy malignant cells over healthy ones. Vesicular stomatitis virus (VSV) is known for its wide range of advantages: a lack of pre-existing immunity, a genome that is easily amenable to manipulation, and rapid growth to high titers in a broad range of cell lines, to name a few. VSV-induced tumor immunity can be enhanced by the delivery of immunostimulatory cytokines. The targeted cytokine delivery to tumors avoids the significant toxicity associated with systemic delivery while also boosting the immune response. To demonstrate this enhanced effect on both tumor growth and survival, a novel recombinant VSV (rVSV)-mIL12-mGMCSF, co-expressing mouse IL-12 (interleukin-12) and GM-CSF (granulocyte-macrophage colony-stimulating factor), was tested alongside rVSV-dM51-GFP (rVSV-GFP) that was injected intratumorally in a syngeneic in vivo C57BL/6 mouse model infused subcutaneously with B16-F10 melanoma cells. The pilot study tested the effect of two viral injections 4 days apart and demonstrated that treatment with the two rVSVs resulted in partial inhibition of tumor growth (TGII of around 40%) and an increased survival rate in animals from the treatment groups. The effect of the two VSVs on immune cell populations will be investigated in future in vivo studies with an optimized experimental design with multiple higher viral doses, as a lack of this information presents a limitation of this study.

The level of transduction efficiency of the target retinal cells affects the choice of AAV serotype and the outcome of gene replacement therapy for inherited retinal diseases. This study focused on the tropism and transduction efficiency of AAV2.7m8-, AAV5-, AAV8-, and AAV9-GFP in ARPE-19 and HEK293 cells. Fluorescence intensity was assessed bi-hourly by means of IncuCyte S3 live imaging microscopy. Within 12 h, AAV2.7m8 demonstrated the highest transduction efficiency at four viral concentrations of 1-, 3-, 6-, and 8 × 10⁴ VG/cell in a dose-dependent manner, followed by AAV5 in ARPE-19 and AAV9 in HEK293 cells. The transduction efficiency of AAV2.7m8 at a dose of 6 × 10⁴ VG/cell was 21, 202, and 323 times higher in ARPE-19 cells and 324, 100, and 52 times higher in HEK293 cells compared to AAV5, AAV8, and AAV9, respectively. This trend remained for 4 days at all viral concentrations, as additionally shown by flow cytometry. At a dose of 6 × 10⁴ VG/cell, AAV2.7m8 (97% GFP-positive cells, GFP +) was nearly two and 10 times as efficient as AAV5 (52% GFP+) and AAV9 or AAV8 (both 9%), respectively, in ARPE-19 cells. In HEK293 cells, 95% of AAV2.7m8-, 26% of AAV9-, 17% of AAV8-, and 12% of AAV5-transduced cells were GFP-positive.

Tea has historically been one of the most popular beverages, and it is currently an economically significant crop cultivated in over 50 countries. The Northwestern Caucasus is one of the northernmost regions for industrial tea cultivation worldwide. The domestication of the tea plant in this region took approximately 150 years, during which plantations spreading from the Ozurgeti region in northern Georgia to the southern city of Maykop in Russia. Consequently, tea plantations in the Northern Caucasus can serve as a source of unique genotypes with exceptional cold tolerance. Tea plants are known to be recalcitrant to Agrobacterium-mediated transfection. Research into optimal transfection and regeneration methodologies, as well as the identification of tea varieties with enhanced transformation efficiency, is an advanced strategy for improving tea plant culture. The aim of this study was to search for the optimal Agrobacterium tumefaciens-mediated transfection protocol for the Kolkhida tea variety. As a result of optimizing the transfection medium with potassium phosphate buffer at the stages of pre-inoculation, inoculation and co-cultivation, the restoration of normal morphology and improvement in the attachment of Agrobacterium cells to the surface of tea explants were observed by scanning electron microscopy. And an effective method of high-efficiency Agrobacteria tumefaciens-mediated transfection of the best local tea cultivar, Kolkhida, was demonstrated for the first time.

INTRODUCTION. The development of adeno-associated virus (AAV)-based gene therapy products in Russia requires establishing reference standards, which are used throughout the pharmaceutical development cycle, and monitoring their stability during the storage period. A preparation of empty capsids of AAV serotype 9 (AAV9) is an appropriate material for a reference standard for empty AAV9 capsids (AAV9 RS).AIM. This study aimed to develop analytical procedures to evaluate the AAV9 RS physicochemical quality parameters for its characterisation and to study its storage stability.MATERIALS AND METHODS. Empty AAV9 capsids were produced in HEK293 suspension culture using serum-free medium and optimised transfection parameters. The next steps involved AAV9 clarification, concentration, and purification by affinity chromatography with AAVx resin and diafiltration. The analysis of AAV9 samples used electrophoresis, transmission electron microscopy, enzyme-linked immunosorbent assay (ELISA), size-exclusion chromatography, dynamic light scattering, spectrophotometry, and bio-layer interferometry. The concentration of capsids was measured by ELISA. Analytical procedures for physical titre determination were developed using an AAV9 standard with a known physical titre. The stability study of the AAV9 RS involved storage at –80 °C for 9 months.RESULTS. Size-exclusion chromatography demonstrated the high purity of the established AAV9 RS, with at least 98% content of the viral capsid monomer. Dynamic light scattering, size-exclusion chromatography, and electron microscopy confirmed that the AAV9 RS was free of aggregates. The stability study showed that the AAV9 RS remained stable for 9 months. Dynamic light scattering and spectrophotometry were deemed optimal methods for routine quality analysis measuring the AAV9 RS physical titre, and bio-layer interferometry was recommended for regular analysis. The viral particle titres determined by these methods ranged from 1.48×10¹³ to 5.6×10¹³.CONCLUSIONS. The AAV9 RS established in this study is suitable for quality control of AAV9-based gene therapy products.