Detection of polyomavirus DNA in human brain tumors

AIM: To investigate surgically resected human brain tumors for the presence of polyomavirus DNA. MATERIALS AND METHODS: 65 samples of surgically resected human brain tumors (57 tumors were of neuroectodermal genesis, 4--mesenchimal, 4--metastatic tumors) were examined for the presence of viral DNA by PCR analysis. RESULTS: DNA of polyomaviruses was found in oligodendroligomas (66.7% of cases), oligodendroastrocytomas (66.7% of cases) and glioblastomas (40% of cases). In metastatic tumors viral infection had not been detected. No correlation between the grade of brain tumors and infection with polyomavirus DNA has been revealed. CONCLUSION: Our data point to importance of investigation of the brain tumors and cerebrospinal fluid for determination of viral infection.     

CRISPR/Cas9-Mediated Targeted DNA Integration: Rearrangements at the Junction of Plant and Plasmid DNA

Targeted DNA integration into known locations in the genome has potential advantages over the random insertional events typically achieved using conventional means of genetic modification. We studied the presence and extent of DNA rearrangements at the junction of plant and transgenic DNA in five lines of Arabidopsis thaliana suspension cells carrying a site-specific integration of target genes. Two types of templates were used to obtain knock-ins, differing in the presence or absence of flanking DNA homologous to the target site in the genome. For the targeted insertion, we selected the region of the histone H3.3 gene with a very high constitutive level of expression. Our studies showed that all five obtained knock-in cell lines have rearrangements at the borders of the integrated sequence. Significant rearrangements, about 100 or more bp from the side of the right flank, were found in all five plant lines. Reorganizations from the left flank at more than 17 bp were found in three out of five lines. The fact that rearrangements were detected for both variants of the knock-in template (with and without flanks) indicates that the presence of flanks does not affect the occurrence of mutations.     

Human Multipotent Mesenchymal Stromal Cell–Derived Extracellular Vesicles Enhance Neuroregeneration in a Rat Model of Sciatic Nerve Crush Injury

Peripheral nerve injury remains a serious problem for medicine, with no effective method of treatment at the moment. The most prominent example of this problem is neonatal brachial plexus palsy, which results from the stretching of the brachial plexus nerves in the birth or perinatal period. Multipotent mesenchymal cells (MSCs) and the extracellular vesicles (EVs) they produce are known to have a marked neuroprotective effect in central nervous system injuries. We suggested that the use of MSCs-derived EVs may be an effective approach to the regeneration of peripheral nerves after injury. Sciatic nerve injury was modeled in rats via crushing, and then a gel containing MSCs–EVs was applied to the injured area. After 15 and 30 days, a histological, physiological, and functional assessment of nerve, dorsal root ganglia (DRG), and innervated muscles’ recovery was performed. Transplantation of EVs to the area of sciatic nerve injury significantly reduced muscle atrophy as compared to the control group. Functional recovery of the innervated muscles, as measured by the extensor postural thrust test, was revealed 30 days after the surgery. We associate the obtained results with EVs-induced neuroprotective mechanisms, which were expressed in a decrease in apoptotic neuronal death and an increase in regeneration-associated proteins NF-200 and GAP-43, as well as in DRG and damaged nerve. We suggest that the therapeutic scheme we used is efficient for the treatment of acute peripheral nervous system injuries and can be transferred to the clinics. However, additional studies are required for a more detailed analysis of neuroprotection mechanisms.     

Synthesis of reactive copolymers with peroxide functionality for cross-linking water-soluble polymers

A series of reactive copolymers with peroxide functionality (RCPFs) were synthesized via radical copolymerization of monomer mixtures in an organic solvent comprised of a peroxide monomer 5-(tert-butyl peroxy)-5-methylhex-1-en-3-yne, acrylamide, maleic anhydride, and butyl methacrylate. Peroxide functionality allows the RCPFs to initiate a variety of radical processes, including cross-linking of organic polymers. Hydrophilic monomer subunits (acrylamide and maleic anhydride) within the RCPF macromolecules promote cross-linking of water-soluble polymers. We aimed to investigate RCPF comonomer ratio and its effects on copolymerization kinetics and composition, as well as physico-chemical and colloidal properties. We also evaluated and characterized the kinetic parameters of the thermal decomposition of peroxide moieties in the synthesized RCPF. Findings revealed that RCPF possessed surface-active properties and reduced surface tension at its aqueous solution–air interface. The data indicated that the decomposition process complied with the first-order kinetics, and complex thermal analysis confirmed the presence of peroxide moieties. RCPFs' ability to cross-link water-soluble polymers was demonstrated on poly(acrylamide) and poly(vinyl alcohol).