Investigation of the Neuroprotective Effect of Halloysite Nanocrystals on the Restorative Function of Mice Brain Neurons

There were studied neuroprotective effects of halloysite nanocrystals on the decrease of protein synthesis – amyloid proteins and activation of neurogenesis in mice with genetic mutation of АРР, PSN -1, PSN – 2 genes. Halloysite inhibits B monoaminoxidase activity, at that A monoaminoxidase activity increases. All studied compounds inhibit Са2+ glutamate-induced hold in synaptosomes. The study was realized on mice of С57BL/6 line with the preliminary reproduction of the defect of beta-amyloid proteins. Nanotubular halloysite was used as a neuroprotector. The analysis of the change of the quantity and localization of aggregates of beta-amyloid proteins before and after using nanotubular halloysite was realized by methods of fluorescent detecting of сonfocal microscopy. There is demonstrated the neuroprotective interaction and stimulation of neurons restoration. The results demonstrated the decrease of beta-amyloid proteins, neurogenesis activation, high antioxidant potential of halloysite nanocrystals that is a precondition of the neuroprotective potential of nanotubular halloysit

Molecular Genetic Analysis of Variability of HA, NA and NP Genes of Influenza Virus А (Compared to H1N1 and H7N9 Strains)

Due to the high antigenic variability and properties for reassortment of influenza A virus genes, analyze the variability of genetic markers of two different antigenic subtypes of influenza A virus strains H1N1 and H7N9 isolated from different hosts (humans and avian). The nucleotide sequences for analysis were taken from the database (NCBI). Using the MEGA6 cluster analysis program and calculating genetic distances using the ClustalW algorithm for the coding sequences of the nucleotides of the HA, NA and NP-protein genes, determinations were made on a sample of influenza strains A. Determination of single nucleotide substitutions at positions with mutations was performed using the Flusurver program. The construction of the dendrogram was carried out using the UPGM group pairwise clustering method, the reliability was calculated using the but-strep analysis. The variability of the neurominidase, hemagglutinin, and nucleoprotein genes was determined by local sequence alignment using the Smith – Waterman algorithm of the VectorNTI-11 program. Consensus sequences (СS) for HA, NA and NP-genes were formed; common conservative areas (motives) were found. The analysis of viral nucleic acids on the variability of the genetic markers of the avian influenza virus HA, NA and NP, coding the virulence factors in the H1N1 and H7N9 subtypes, showed genetic variability (variability) of the hemagglutinin and neurominidase genes. A sample of gene sequences showed that the HA genes of the avian influenza virus have more interstitial polymorphism than the NA and NP protein genes. Genetic markers of high variability are the H1N1 subtype hemagglutinin genes and the NA genes in the H7N9 subtype. The article analyzes the structural features of the genes of surface proteins and nucleoproteins of influenza viruses AH1N1 and AH7N9. A certain degree of synonymity of nucleotide substitutions is determined. The relationship between the distribution of nucleotide polymorphism and indicators of synonymous and non-synonymous substitutions is established. The high variability of the NA gene, and somewhat less NA, determines the ability of the avian influenza virus, in particular its highly virulent strain H1N1 and less virulent H7N9, to overcome the interspecific barrier, whereas the replication factor encoded by the NP gene is less important for overcoming the interspecies barrier, which causes its low compared with ON and NA variability

Development of Anexpress-method for Influence and Genotyping of H1N1 and H7N9 Virus Avian Influenza a Strains by PCR-RFLP Analysis

Epizootic monitoring in recent years suggests that the highly pathogenic avian influenza A virus (H1N1) and (H7N9) actively circulate in the Eurasian countries. By 2016 - 2019 1.6 thousand outbreaks were recorded. For 2016 - 2019, 1.6 thousand cases of outbreaks were recorded. Of these, there are 872 cases in Europe. The monitoring of infected birds, both migratory and poultry, in places of cross-contact in Ukraine is relevant for preventing outbreaks of epizooties.The aim of the study. To develop an express method for the identification and determination of bird flu virus A H1N1 and H7N9 strains, based on a polymerase chain reaction with analysis of restriction fragment length polymorphism (PCR-RFLP) of the virus RNA.Results and discussion. The in silico analysis of the HA, NA, and NP gene amplicons allowed in silico to calculate the primers to the variable loci of the investigated genes, to calculate the reaction conditions, to determine restriction sites for the restriction enzyme to obtain theoretical PCR electrophoregrams. An express method for the detection and identification of influenza A H1N1 and H7N9 virus by three genes (HA, NA, and NP) of H1N1 and H7N9 RNA in polymerase chain reaction, combined with RFLP analysis, was developed. The method of rapid diagnostics is able to detect avian influenza virus A H1N1 and H7N9 and differentiate it from samples of other pathogens of viral infections of birds and animals. It was established, that the PCR-RFLP rapid diagnostic method is able to detect influenza A virus RNA of H1N1 and H7N9 strains with high sensitivity (100 % sensitivity).Conclusions.The developed method of PCR-based rapid identification, combined with RFLP analysis, makes it possible to significantly simplify the method of identification due to specific amplification of an RNA region having a polymorphic restriction site. Testing of this locus is possible by pre-PCR and restriction of the amplified fragment. The method of express - diagnosis of PLR-RFLP has been established for detecting RNA virus influenza A of high pathogenic H1N1 and H7N9 strains with high indicators of sensitivity (100 % sensitivity